Authorship：Last author,　Corresponding author   Language：Others  

DOI： 10.1101/2023.11.27.568629

CiNii Research

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Language：Others   Publisher：Cold Spring Harbor Laboratory  

Cell states are regulated by the response of signaling pathways to receptor ligand-binding and inter-cellular interactions. High-resolution imaging has been attempted to explore the dynamics of these processes and, recently, multiplexed imaging has profiled cell states by achieving comprehensive acquisition of spatial protein information from cells. However, resolution is still compromised when visualizing activated signals. Here we developed Precise Emission Canceling Antibody (PECAb) that have cleavable fluorescent labeling. PECAbs allow high-resolution sequential imaging using hundreds of antibodies and allow reconstruction of the spatiotemporal dynamics of signaling pathways. Additionally, combining this approach with seq-smFISH can effectively classify cells and identify their signal activation states in human tissue. Overall, the PECAb system can serve as a comprehensive platform for analyzing complex cell processes.

DOI： 10.1101/2023.10.17.561810

CiNii Research

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Authorship：Lead author,　Corresponding author   Language：English   Publisher：Elsevier {BV}  

Higher-order genomic structures play a critical role in regulating gene expression by influencing the spatial proximity of promoters and enhancers. Live-cell imaging studies have demonstrated that three-dimensional genome structures undergo dynamic changes over time. Transcription is also dynamic, with genes frequently switching between active and inactive states. Recent observations suggest that the formation of condensates, composed of transcription-related factors, RNA, and RNA-binding proteins, around genes can regulate transcription. Advancements in technology have facilitated the visualization of the intricate spatiotemporal relationship between higher-order genomic structures, condensate formation, and transcriptional activity in living cells.

DOI： 10.1016/j.sbi.2023.102615

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Authorship：Last author,　Corresponding author   Language：Others   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media {LLC}  

STREAMING-tag system reveals spatiotemporal relationships between transcriptional regulatory factors and transcriptional activity

DOI： 10.1038/s41467-022-35286-2

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Life Science Alliance  

Ribosomal protein L5 facilitates rDNA-bundled condensate and nucleolar assembly
The nucleolus is the site of ribosome assembly and formed through liquid-liquid phase separation. Multiple ribosomal DNA (rDNA) arrays are bundled in the nucleolus, but the underlying mechanism and significance are unknown. In the present study, we performed high-content screening followed by image profiling with the wndchrm machine learning algorithm. We revealed that cells lacking a specific 60S ribosomal protein set exhibited common nucleolar disintegration. The depletion of RPL5 (also known as uL18), the liquid-liquid phase separation facilitator, was most effective, and resulted in an enlarged and un-separated sub-nucleolar compartment. Single-molecule tracking analysis revealed less-constrained mobility of its components. rDNA arrays were also unbundled. These results were recapitulated by a coarse-grained molecular dynamics model. Transcription and processing of ribosomal RNA were repressed in these aberrant nucleoli. Consistently, the nucleoli were disordered in peripheral blood cells from a Diamond-Blackfan anemia patient harboring a heterozygous, large deletion in RPL5 Our combinatorial analyses newly define the role of RPL5 in rDNA array bundling and the biophysical properties of the nucleolus, which may contribute to the etiology of ribosomopathy.

DOI： 10.26508/lsa.202101045

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PLoS ONE  

iPSC reprogramming-mediated aneuploidy correction in autosomal trisomy syndromes
Trisomy 21, 18, and 13 are the major autosomal aneuploidy disorders in humans. They are mostly derived from chromosome non-disjunction in maternal meiosis, and the extra trisomic chromosome can cause several congenital malformations. Various genes on the trisomic chromosomes are intricately involved in the development of disease, and fundamental treatments have not yet been established. However, chromosome therapy has been developed to correct the extra chromosome in cultured patient cells, and it was recently reported that during reprogramming into iPSCs, fibroblasts from a Down syndrome patient lost the extra chromosome 21 due to a phenomenon called trisomy-biased chromosome loss. To gain preliminary insights into the underlying mechanism of trisomy rescue during the early stages of reprogramming, we reprogrammed skin fibroblasts from patients with trisomy syndromes 21, 18, 13, and 9 to iPSC, and evaluated the genomes of the individual iPSC colonies by molecular cytogenetic techniques. We report the spontaneous correction from trisomy to disomy upon cell reprogramming in at least one cell line examined from each of the trisomy syndromes, and three possible combinations of chromosomes were selected in the isogenic trisomy-rescued iPSC clones. Single nucleotide polymorphism analysis showed that the trisomy-rescued clones exhibited either heterodisomy or segmental uniparental isodisomy, ruling out the possibility that two trisomic chromosomes were lost simultaneously and the remaining one was duplicated, suggesting instead that one trisomic chromosome was lost to generate disomic cells. These results demonstrated that trisomy rescue may be a phenomenon with random loss of the extra chromosome and subsequent selection for disomic iPSCs, which is analogous to the karyotype correction in early preimplantation embryos. Our finding is relevant for elucidating the mechanisms of autonomous karyotype correction and future application in basic and clinical research on aneuploidy disorders.

DOI： 10.1371/journal.pone.0264965

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Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/s41594-020-0493-6

Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

Transcriptional bursting is the stochastic activation and inactivation of promoters, contributing to cell-to-cell heterogeneity in gene expression. However, the mechanism underlying the regulation of transcriptional bursting kinetics (burst size and frequency) in mammalian cells remains elusive. In this study, we performed single-cell RNA sequencing to analyze the intrinsic noise and mRNA levels for elucidating the transcriptional bursting kinetics in mouse embryonic stem cells. Informatics analyses and functional assays revealed that transcriptional bursting kinetics was regulated by a combination of promoter- and gene body-binding proteins, including the polycomb repressive complex 2 and transcription elongation factors. Furthermore, large-scale CRISPR-Cas9-based screening identified that the Akt/MAPK signaling pathway regulated bursting kinetics by modulating transcription elongation efficiency. These results uncovered the key molecular mechanisms underlying transcriptional bursting and cell-to-cell gene expression noise in mammalian cells.

DOI： 10.1126/sciadv.aaz6699

Language：English   Publishing type：Research paper (scientific journal)  

Insufficiency of ciliary cholesterol in hereditary Zellweger syndrome

DOI： 10.15252/embj.2019103499

Language：English   Publishing type：Research paper (scientific journal)  

PHi-C: deciphering Hi-C data into polymer dynamics
<title>Abstract</title>
Genomes are spatiotemporally organized within the cell nucleus. Genome-wide chromosome conformation capture (Hi-C) technologies have uncovered the 3D genome organization. Furthermore, live-cell imaging experiments have revealed that genomes are functional in 4D. Although computational modeling methods can convert 2D Hi-C data into population-averaged static 3D genome models, exploring 4D genome nature based on 2D Hi-C data remains lacking. Here, we describe a 4D simulation method, PHi-C (polymer dynamics deciphered from Hi-C data), that depicts 4D genome features from 2D Hi-C data by polymer modeling. PHi-C allows users to interpret 2D Hi-C data as physical interaction parameters within single chromosomes. The physical interaction parameters can then be used in the simulations and analyses to demonstrate dynamic characteristics of genomic loci and chromosomes as observed in live-cell imaging experiments. PHi-C is available at https://github.com/soyashinkai/PHi-C.

DOI： 10.1093/nargab/lqaa020

Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1371/journal.pcbi.1007289

Language：English   Publishing type：Research paper (scientific journal)  

Single-Molecule Nanoscopy Elucidates RNA Polymerase II Transcription at Single Genes in Live Cells

DOI： 10.1016/j.cell.2019.05.029

Language：English   Publishing type：Research paper (scientific journal)  

The nuclear positioning and chromatin dynamics of eukaryotic genes are closely related to the regulation of gene expression, but they have not been well examined during early development, which is accompanied by rapid cell cycle progression and dynamic changes in nuclear organization, such as nuclear size and chromatin constitution. In this study, we focused on the early development of the sea urchin Hemicentrotus pulcherrimus and performed three-dimensional fluorescence in situ hybridization of gene loci encoding early histones (one of the types of histone in sea urchin). There are two non-allelic early histone gene loci per sea urchin genome. We found that during the morula stage, when the early histone gene expression levels are at their maximum, interchromosomal interactions were often formed between the early histone gene loci on separate chromosomes and that the gene loci were directed to locate to more interior positions. Furthermore, these interactions were associated with the active transcription of the early histone genes. Thus, such dynamic interchromosomal interactions may contribute to the efficient synthesis of early histone mRNA during the morula stage of sea urchin development.

DOI： 10.1242/jcs.206862

Language：English   Publishing type：Research paper (scientific journal)  

Ligation-mediated polymerase chain reaction (LM-PCR) is a common technique for amplification of a pool of DNA fragments. Here, a double-stranded oligonucleotide consisting of two primer sequences in back-to-back orientation was designed as an adapter for LM-PCR. When DNA fragments were ligated with this adapter, the fragments were sandwiched between two adapters in random orientations. In the ensuing PCR, ligation products linked at each end to an opposite side of the adapter, i.e. to a distinct primer sequence, were preferentially amplified compared with products linked at each end to an identical primer sequence. The use of this adapter in LM-PCR reduced the impairment of PCR by substrate DNA with a high GC content, compared with the use of traditional LM-PCR adapters. This result suggested that our method has the potential to contribute to reduction of the amplification bias that is caused by an intrinsic property of the sequence context in substrate DNA. A DNA preparation obtained from a chromatin immunoprecipitation assay using pulldown of a specific form of histone H3 was successfully amplified using the modified LM-PCR, and the amplified products could be used as probes in a fluorescence in situ hybridization analysis. (C) 2017 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ab.2017.05.011

Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

The relationship between genome organization and gene expression has recently been established. However, the relationships between spatial organization, dynamics, and transcriptional regulation of the genome remain unknown. In this study, we developed a live-imaging method for simultaneous measurements of the transcriptional activity and nuclear position of endogenous genes, which we termed the 'Real-time Observation of Localization and EXpression (ROLEX)' system. We demonstrated that ROLEX is highly specific and does not affect the expression level of the target gene. ROLEX enabled detection of sub-genome-wide mobility changes that depended on the state of Nanog transactivation in embryonic stem cells. We believe that the ROLEX system will become a powerful tool for exploring the relationship between transcription and nuclear dynamics in living cells.

DOI： 10.1093/nar/gkv624

Language：English   Publishing type：Research paper (scientific journal)  

The primary cilium is an antenna-like, microtubule-based organelle on the surface of most vertebrate cells for receiving extracellular information. Although primary cilia form in the quiescent phase, ciliary disassembly occurs when quiescent cells re-enter the proliferative phase. It was shown that a mitotic kinase, Polo-like kinase 1 (PLK1), is required for cell-proliferation-coupled primary cilia disassembly. Here, we report that kinesin superfamily protein 2A (KIF2A), phosphorylated at T554 by PLK1, exhibits microtubule-depolymerizing activity at the mother centriole to disassemble the primary cilium in a growth-signal-dependent manner. KIF2A-deficient hTERT-RPE1 cells showed the impairment of primary cilia disassembly following growth stimulation. It was also found that the PLK1-KIF2A pathway is constitutively active in cells from patients with premature chromatid separation (PCS) syndrome and is responsible for defective ciliogenesis in this syndrome. These findings provide insights into the roles of the PLK1-KIF2A pathway in physiological cilia disassembly and cilia-associated disorders.

DOI： 10.1016/j.celrep.2015.01.003

Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

Mouse embryonic stem cells (mESCs) are self-renewing and capable of differentiating into any of the three germ layers. An interesting feature of mESCs is the presence of cell-to-cell heterogeneity in gene expression that may be responsible for cell fate decisions. Nanog, a key transcription factor for pluripotency, displays heterogeneous expression in mESCs, via mechanisms that are not fully understood. To understand this variability, we quantitatively analyzed Nanog transcription and found that Nanog was both infrequently transcribed, and transcribed in a pulsatile and stochastic manner. It is possible that such stochastic transcriptional activation could contribute to the heterogeneity observed in Nanog expression as "intrinsic noise.'' To discriminate the effects of both intrinsic noise from other (extrinsic) noise on the expression variability of Nanog mRNA, we performed allele-specific single-molecule RNA fluorescent in situ hybridization in a reporter cell line and found that intrinsic noise contributed to approximately 45% of the total variability in Nanog expression. Furthermore, we found that Nanog mRNA and protein levels were well correlated in individual cells. These results suggest that stochastic promoter activation significantly affects the Nanog expression variability in mESCs.

DOI： 10.1038/srep07125

Repository Public URL： https://hdl.handle.net/2324/7237095

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

Cancer-prone syndrome of premature chromatid separation with mosaic variegated aneuploidy [PCS (MVA) syndrome] is a rare autosomal recessive disorder characterized by constitutional aneuploidy and a high risk of childhood cancer. We previously reported monoallelicmutations in the BUB1B gene (encoding BUBR1) in seven Japanese families with the syndrome. No second mutation was found in the opposite allele of any of the families studied, although a conserved BUB1B haplotype and a decreased transcript were identified. To clarify the molecular pathology of the second allele, we extended our mutational search to a candidate region surrounding BUB1B. A unique single nucleotide substitution, G > A at ss802470619, was identified in an intergenic region 44 kb upstream of a BUB1B transcription start site, which cosegregated with the disorder. To examine whether this is the causal mutation, we designed a transcription activator-like effector nuclease-mediated two-step single-base pair editing strategy and biallelically introduced this substitution into cultured human cells. The cell clones showed reduced BUB1B transcripts, increased PCS frequency, and MVA, which are the hallmarks of the syndrome. We also encountered a case of a Japanese infant with PCS (MVA) syndrome carrying a homozygous single nucleotide substitution at ss802470619. These results suggested that the nucleotide substitution identified was the causal mutation of PCS (MVA) syndrome.

DOI： 10.1073/pnas.1317008111

Language：English   Publishing type：Research paper (scientific journal)  

Transcription activator-like effector (TALE) nuclease (TALEN) is a site-specific nuclease, which can be freely designed and easily constructed. Numerous methods of constructing TALENs harboring different TALE scaffolds and repeat variants have recently been reported. However, the functionalities of structurally different TALENs have not yet been compared. Here, we report on the functional differences among several types of TALENs targeting the same loci. Using HEK293T cell-based single-strand annealing and Cel-I nuclease assays, we found that TALENs with periodically-patterned repeat variants harboring non-repeat-variable di-residue (non-RVD) variations (Platinum TALENs) showed higher activities than TALENs without non-RVD variations. Furthermore, the efficiencies of gene disruption mediated by Platinum TALENs in frogs and rats were significantly higher than in previous reports. This study therefore demonstrated an efficient system for the construction of these highly active Platinum TALENs (Platinum Gate system), which could establish a new standard in TALEN engineering.

DOI： 10.1038/srep03379

Language：English   Publishing type：Research paper (scientific journal)  

Recently, gene editing with transcription activator-like effector nucleases (TALENs) has been used in the life sciences. TALENs can be easily customized to recognize a specific DNA sequence and efficiently introduce double-strand breaks at the targeted genomic locus. Subsequent non-homologous end-joining repair leads to targeted gene disruption by base insertion, deletion, or both. Here, to readily evaluate the efficacy of TALENs in Xenopus laevis embryos, we performed the targeted gene disruption of tyrosinase (tyr) and pax6 genes that are involved in pigmentation and eye formation, respectively. We constructed TALENs targeting tyr and pax6 and injected their mRNAs into fertilized eggs at the one-cell stage. Expectedly, introduction of tyr TALEN mRNA resulted in drastic loss of pigmentation with high efficiency. Similarly, for pax6, TALENs led to deformed eyes in the injected embryos. We confirmed mutations of the target alleles by restriction enzyme digestion and sequence analyses of genomic PCR products. Surprisingly, not only biallelic but also paralogous, gene disruption was observed. Our results demonstrate that targeted gene disruption by TALENs provides a method comparable to antisense morpholinos in analyzing gene function in Xenopus F0 embryos, but also applies beyond embryogenesis to any life stage. (C) 2013. Published by The Company of Biologists Ltd.

DOI： 10.1242/bio.20133855

Language：English   Publishing type：Research paper (scientific journal)  

Transcription activator-like effector nucleases (TALENs) have recently arisen as effective tools for targeted genome engineering. Here, we report streamlined methods for the construction and evaluation of TALENs based on the 'Golden Gate TALEN and TAL Effector Kit' (Addgene). We diminished array vector requirements and increased assembly rates using six-module concatemerization. We altered the architecture of the native TALEN protein to increase nuclease activity and replaced the final destination vector with a mammalian expression/in vitro transcription vector bearing both CMV and T7 promoters. Using our methods, the whole process, from initiating construction to completing evaluation directly in mammalian cells, requires only 1 week. Furthermore, TALENs constructed in this manner may be directly applied to transfection of cultured cells or mRNA synthesis for use in animals and embryos. In this article, we show genomic modification of HEK293T cells, human induced pluripotent stem cells, Drosophila melanogaster, Danio rerio and Xenopus laevis, using custom-made TALENs constructed and evaluated with our protocol. Our methods are more time efficient compared with conventional yeast-based evaluation methods and provide a more accessible and effective protocol for the application of TALENs in various model organisms. © 2013 The Authors Genes to Cells © 2013 by the Molecular Biology Society of Japan and Wiley Publishing Asia Pty Ltd.

DOI： 10.1111/gtc.12037

Language：English   Publishing type：Research paper (scientific journal)  

Lens growth involves the proliferation of epithelial cells, followed by their migration to the equator region and differentiation into secondary fiber cells. It is widely accepted that fibroblast growth factor (FGF) signaling is required for the differentiation of lens epithelial cells into crystallin-rich fibers, but this signaling is insufficient to induce full differentiation. To better understand lens development, investigatory and functional analyses of novel molecules are required. Here, we demonstrate that Equarin, which is a novel secreted molecule, was expressed exclusively in the lens equator region during chick lens development. Equarin upregulated the expression of fiber markers, as demonstrated using in ovo electroporation. In a primary lens cell culture, Equarin promoted the biochemical and morphological changes associated with the differentiation of lens epithelial cells to fibers. A loss-of-function analysis was performed using zinc-finger nucleases targeting the Equarin gene. Lens cell differentiation was markedly inhibited when endogenous Equarin was blocked, indicating that Equarin was essential for normal chick lens differentiation. Furthermore, biochemical analysis showed that Equarin directly bound to FGFs and heparan sulfate proteoglycan and thereby upregulated the expression of phospho-ERK1/2 (ERK-P) proteins, the downstream of the FGF signaling pathway, in vivo and in vitro. Conversely, the absence of endogenous Equarin clearly diminished FGF-induced fiber differentiation. Taken together, our results suggest that Equarin is involved as an FGF modulator in chick lens differentiation. (C) 2012 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ydbio.2012.05.029

Language：English   Publishing type：Research paper (scientific journal)  

Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically relatively basal and comprise many pests. However, the absence of a sophisticated genetic model system, or targeted gene-manipulation system, has limited research on hemimetabolous species. Here we use zinc-finger nuclease and transcription activator-like effector nuclease technologies to produce genetic knockouts in the hemimetabolous insect Gryllus bimaculatus. Following the microinjection of mRNAs encoding zinc-finger nucleases or transcription activator-like effector nucleases into cricket embryos, targeting of a transgene or endogenous gene results in sequence-specific mutations. Up to 48% of founder animals transmit disrupted gene alleles after zinc-finger nucleases microinjection compared with 17% after microinjection of transcription activator-like effector nucleases. Heterozygous offspring is selected using mutation detection assays that use a Surveyor (Cel-I) nuclease, and subsequent sibling crosses create homozygous knockout crickets. This approach is independent from a mutant phenotype or the genetic tractability of the organism of interest and can potentially be applied to manage insect pests using a non-transgenic strategy.

DOI： 10.1038/ncomms2020

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

To understand complex biological systems, such as the development of multicellular organisms, it is important to characterize the gene expression dynamics. However, there is currently no universal technique for targeted insertion of reporter genes and quantitative imaging in multicellular model systems. Recently, genome editing using zinc-finger nucleases (ZFNs) has been reported in several models. ZFNs consist of a zinc-finger DNA-binding array with the nuclease domain of the restriction enzyme FokI and facilitate targeted transgene insertion. In this study, we successfully inserted a GFP reporter cassette into the HpEts1 gene locus of the sea urchin, Hemicentrotus pulcherrimus. We achieved this insertion by injecting eggs with a pair of ZFNs for HpEts1 with a targeting donor construct that contained similar to 1-kb homology arms and a 2A-histone H2B-GFP cassette. We increased the efficiency of the ZFN-mediated targeted transgene insertion by in situ linearization of the targeting donor construct and cointroduction of an mRNA for a dominant-negative form of HpLig4, which encodes the H. pulcherrimus homolog of DNA ligase IV required for error-prone nonhomologous end joining. We measured the fluorescence intensity of GFP at the single-cell level in living embryos during development and found that there was variation in HpEts1 expression among the primary mesenchyme cells. These findings demonstrate the feasibility of ZFN-mediated targeted transgene insertion to enable quantification of the expression levels of endogenous genes during development in living sea urchin embryos.

DOI： 10.1073/pnas.1202768109

Language：English   Publishing type：Research paper (scientific journal)  

Zinc-finger nucleases (ZFNs) are engineered nucleases that induce DNA double-strand breaks (DSBs) at target sequences. They have been used as tools for generating targeted mutations in the genomes of multiple organisms in both animals and plants. The DSB induced by ZFNs is repaired by non-homologous end joining (NHEJ) or by homologous recombination (HR) mechanisms. Non-homologous end joining induces some errors because it is independent of a reference DNA sequence. Through the NHEJ mechanism, ZFNs generate insertional or deletional mutations at the target sequence. We examined the usability, specificity and toxicity of ZFNs in the basal chordate Ciona intestinalis. As the target of ZFNs, we chose an enhanced green fluorescent protein (EGFP) gene artificially inserted in the C. intestinalis genome because this locus is neutral for the development and growth of C. intestinalis, and the efficiency of mutagenesis with ZFNs can thus be determined without any bias. We introduced EGFP-ZFN mRNAs into the embryos of an EGFP-transgenic line and observed the mutation frequency in the target site of EGFP . We also examined the effects of the EGFP-ZFNs at off-target sites resembling the EGFP target sequence in the C. intestinalis genome in order to examine the specificity of ZFNs. We further investigated the influence of ZFNs on embryogenesis, and showed that adequate amounts of ZFNs, which do not disrupt embryogenesis, can efficiently induce mutations on the on-target site with less effect on the off-target sites. This suggests that target mutagenesis with ZFNs will be a powerful technique in C. intestinalis.

DOI： 10.1111/j.1440-169X.2012.01355.x

Language：English   Publishing type：Research paper (scientific journal)  

Zinc-finger nucleases (ZFNs) are artificial enzymes that create site-specific double-strand breaks and thereby induce targeted genome editing. Here, we demonstrated successful gene disruption in somatic and germ cells of medaka (Oryzias latipes) using ZFN to target exogenous EGFP genes. Embryos that were injected with an RNA sequence pair coding for ZFNs showed mosaic loss of green fluorescent protein fluorescence in skeletal muscle. A number of mutations that included both deletions and insertions were identified within the ZFN target site in each embryo, whereas no mutations were found at the non-targeted sites. In addition, ZFN-induced mutations were introduced in germ cells and efficiently transmitted to the next generation. The mutation frequency varied (6100%) in the germ cells from each founder, and a founder carried more than two types of mutation in germ cells. Our results have introduced the possibility of targeted gene disruption and reverse genetics in medaka.

DOI： 10.1111/j.1440-169X.2012.01357.x

Language：English   Publishing type：Research paper (scientific journal)  

Sulfatases such as arylsulfatase and heparan sulfate 6-O-endosulfatase play important roles in morphogenesis during sea urchin development. For the activation of these sulfatases, C alpha-formylglycine formation by sulfatase modifying factor (Sumf) is required. In this study, to clarify the regulatory mechanisms for the activation of sulfatases during sea urchin development, we examined the expression and function of the Hemicentrotus pulcherrimus homologs of Sumf1 and Sumf2 (HpSumf1 and HpSumf2, respectively). Expression of HpSumf1 but not HpSumf2 mRNA was dynamically changed during early development. Functional analyses of recombinant HpSumf1 and HpSumf2 using HEK293T cells expressing mouse arylsulfatase A (ArsA) indicated that HpSumf1 and HpSumf2 were both able to activate mammalian ArsA. Knockdown of HpSumf1 using morpholino antisense oligonucleotides caused abnormal spicule formation in the sea urchin embryo. Injection of HpSumf2 mRNA had no effect on skeletogenesis, while injection of HpSumf1 mRNA induced severe supernumerary spicule formation. Taken together, these findings suggest that HpSumf1 is involved in the activation of sulfatases required for control of skeletogenesis.

DOI： 10.1007/s00427-011-0368-2

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

We showed that engineered zinc-finger nucleases (ZFNs), which consist of a zinc-finger DNA-binding array and a nuclease domain of the restriction enzyme FokI, can introduce mutations at a specific genomic site in the sea urchin embryo. Using bacterial one-hybrid screening with zinc-finger randomized libraries and a single-strand annealing assay in cultured cells, ZFNs targeting the sea urchin Hemicentrotus pulcherrimus homologue of HesC (HpHesC) were efficiently selected. Consistent with the phenotype observed in embryos injected with an antisense morpholino oligonucleotide against HpHesC, an increase in the primary mesenchyme cell population was observed in embryos injected with a pair of HpHesC ZFN mRNAs. In addition, sequence analysis of the mutations showed that deletions and insertions occurred at the HpHesC target site in the embryos injected with the HpHesC ZFN mRNAs. These results suggest that targeted gene disruption using ZFNs is feasible for the sea urchin embryo.

DOI： 10.1111/j.1365-2443.2010.01425.x

Language：English   Publishing type：Research paper (scientific journal)  

MicroRNAs are single-stranded RNA molecules with a length of 19-25 nucleotides, which play roles in various biological phenomena, including development, differentiation, apoptosis, by regulating target gene expression. Although the presence of microRNA molecules in sea urchin and the expression of genes involved in microRNA biogenesis during sea urchin development have been reported recently, the function of microRNA in sea urchin development remains to be elucidated. In this study, to understand the function of microRNA in the early development of sea urchin, we focused on Dicer, an essential enzyme for biosynthesis of mature microRNA. We determined the nucleotide sequence of cDNA for a Dicer homolog in the sea urchin, Hemicentrotus pulcherrimus, HpDcr, and found that functional domains of Dicer proteins are conserved in HpDcr. Analyses of its pattern of expression showed that HpDcr mRNA is expressed in embryos at all developmental stages analyzed, and seems to distribute asymmetrically at the morula and later stages. Knockdown of HpDcr resulted in anomalous morphogenesis, such as impairment of gastrulation and skeletogenesis at the mesenchyme blastula stage and later stages, and alteration of mRNA levels of cell type-specific genes. Thus, HpDcr plays important roles in morphogenesis in sea urchin embryos, suggesting that miRNA could be involved in the early development of sea urchin by regulating target gene expression.

DOI： 10.2108/zsj.27.477

Language：English   Publishing type：Research paper (scientific journal)  

The nanos genes play important roles in the development of primordial germ cells in animal species. In the sea urchin, Hemicentrotus pulcherrimus, small micromere descendants specifically express HpNanos mRNA and this expression continues in the left coelomic pouch, which produces the major component of the adult rudiment. In this study, we showed that morpholino knockdown of HpNanos resulted in a delay of primary mesenchyme cell ingression and a decrease in the number of cells comprising the left coelomic pouch. Knockdown analysis in chimeras and whole embryos revealed the disappearance of small micromere descendants from the archenteron tip. Furthermore, the expression of HpNanos mRNA was induced in other cell lineages in the HpNanos-knockdown and micromere-deleted embryos. Taken together, our results suggest that HpNanos is involved in the inductive interaction of small micromere descendants with other cell lineages, and that HpNanos is required for the survival of small micromere descendants. Developmental Dynamics 238:2511-2521, 2009. (c) 2009 Wiley-Liss, Inc.

DOI： 10.1002/dvdy.22074

Language：English   Publishing type：Research paper (scientific journal)  

In sea urchin embryos, Notch signaling is required to segregate non-skeletogenic mesoderm from early endomesoderm, and is involved in endoderm development. To further investigate the role of Notch signaling in the endoderm cell lineage, we cloned a cDNA for the Hemicentrotus pulcherrimus ortholog of Suppressor of Hairless (HpSu(H)), which is a major mediator of the Notch signaling pathway, examined the expression during development and performed a functional analysis. HpSu(H) mRNA was ubiquitously expressed up to the unhatched blastula stage, and expression was exclusively detected in the vegetal plate region from the hatched blastula stage and then in the archenteron at the gastrula stage. Perturbation of HpSu(H) by injection of the dominant negative form of HpSu(H) (dn-HpSu(H)) mRNA into fertilized eggs led to the disappearance of secondary mesenchyme cells at the tip of the archenteron in the gastrula and pigment cells in the pluteus larva, confirming that Notch signaling is required for non-skeletogenic mesoderm specification. In addition, injection of relatively high amounts of dn-HpSu(H) mRNA caused a defect or atrophy of the foregut in the archenteron at the pluteus stage. This result strongly suggests that Notch signaling is involved in foregut development during sea urchin development.

DOI： 10.2108/zsj.26.686

Language：English   Publishing type：Research paper (scientific journal)  

For the efficient generation of transgenic sea urchins, we have adopted an I-SceI meganuclease-mediated transgenesis method. Several types of promoter-GFP gene constructs flanked by two I-SceI recognition sequences were co-injected with I-SceI into sea urchin fertilized eggs. Using cell-lineage-specific promoter constructs, the frequency of transgene expression was elevated, and their level of mozaicism was reduced. The addition of the Ars insulator sequence, which is known to block the enhancer activity and protect transgenes from position effects, led to a reduction in ectopic transgene expression and an elevation of transgene expression frequency in this I-SceI-mediated system. However, the magnitude of the effects of the Ars insulator was dependent upon the promoter constructs. QPCR analysis also showed that the Ars insulator increases the transgene copy number. These results suggest that the I-SceI-mediated method using the Ars insulator is advantageous for transgenesis in the sea urchin embryo.

DOI： 10.1002/dvdy.21690

Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

In sea urchin development, micromere descendants play important roles in skeletogenesis and induction of gastrulation. We previously reported that the T-brain homolog of sea urchin Hemicentrotus pulcherrimus, HpTb expresses specifically in micromere descendants and is required for induction of gastrulation and skeletogenesis. Thus, HpTb is thought to play important roles in the function of micromere-lineage cells. To identify cis-regulatory regions responsible for spatio-temporal gene expression of HpTb, we isolated similar to 7 kb genomic region of HpTb gene and showed that GFP expression driven by this region exhibits the spatio-temporal pattern corresponding substantially to that of endogenous HpTb expression. Deletion of interspecifically conserved C2 and C4 regions resulted in an increase of ectopic expression. Mutations in Hairy family and Snail family consensus sequences in C1 and C2 regions also increased ectopic expression. Furthermore, we demonstrated that C4 region functions as enhancer, and that three Ets family consensus sequences are involved in this activity but not in spatial regulation. Therefore, we concluded that expression of HpTb gene is regulated by multiple cis-regulatory elements. (c) 2007 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.mod.2007.10.009

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Proceedings of the National Academy of Sciences  

Intercellular transmission of messenger RNA (mRNA) is being explored in mammalian species using immortal cell lines. Here, we uncover an intercellular mRNA transfer phenomenon that allows for the adaptation and reprogramming of human primed pluripotent stem cells (hPSCs). This process is induced by the direct cell contact-mediated coculture with mouse embryonic stem cells under the condition impermissible for primed hPSC culture. Mouse-derived mRNA contents are transmitted into adapted hPSCs only in the coculture. Transfer-specific mRNA analysis shows the enrichment for divergent biological pathways involving transcription/translational machinery and stress-coping mechanisms, wherein such transfer is diminished when direct cell contacts are lost. After 5 d of coculture with mouse embryonic stem cells, surface marker analysis and global gene profiling confirmed that mRNA transfer-prone hPSC efficiently gains a naïve-like state. Furthermore, transfer-specific knockdown experiments targeting mouse-specific transcription factor-coding mRNAs in hPSC show that mouse-derived Tfcp2l1 , Tfap2c, and Klf4 are indispensable for human naïve-like conversion. Thus, interspecies mRNA transfer triggers cellular reprogramming in mammalian cells. Our results support that episodic mRNA transfer can occur in cell cooperative and competitive processes, which provides a fresh perspective on understanding the roles of mRNA mobility for intra- and interspecies cellular communications.

DOI： 10.1073/pnas.2413351122

Web of Science

Scopus

PubMed

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Publisher：Cold Spring Harbor Laboratory  

ABSTRACT

Spatial omics enables comprehensive mapping of cell types and states in their spatial context, providing profound insights into cellular communication and tissue organization. However, analyzing large tissue sections, especially crucial for clinical applications, remains a significant challenge due to the computational demands of current image processing methods. To overcome these limitations, we developed MEGA-FISH, a flexible, GPU-accelerated Python framework optimized for large-scale spatial omics image analysis. Benchmarking on simulated and tissue images demonstrated that MEGA-FISH achieved high accuracy in spot detection while significantly reducing processing times compared with established tools. The framework’s adaptable computational capabilities optimize resource allocation (e.g., GPU or multi-core CPU) for diverse tasks, and its scalable architecture enables integration with advanced imaging and segmentation techniques. By bridging cutting-edge imaging methods and single-cell analysis, MEGA-FISH provides an efficient platform for multi-modal analysis and advances research and clinical applications of spatial omics at organ and organism scales.

DOI： 10.1101/2024.12.04.626913

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Publisher：Cold Spring Harbor Laboratory  

ABSTRACT

In eukaryotic genomes, regulated access and communication between cis-regulatory elements (CREs) are necessary for enhancer-mediated transcription of genes. The molecular framework of the chromatin organization underlying such communication remains poorly understood. To better understand it, we develop a multiscale modeling pipeline to build near-atomistic models of the 200 kbNanoggene locus in mouse embryonic stem cells comprising nucleosomes, transcription factors, co-activators, and RNA polymerase II-Mediator complexes. By integrating diverse experimental data, including protein localization, genomic interaction frequencies, cryo-electron microscopy, and single-molecule fluorescence studies, our model offers novel insights into chromatin organization and its role in enhancer-promoter communication. The models equilibrated by high-performance molecular dynamics simulations span a scale of ∼350 nm, revealing an experimentally consistent local and global organization of chromatin and transcriptional machinery. Our models elucidate that the sequence-regulated chromatin accessibility facilitates the recruitment of transcription regulatory proteins exclusively at CREs, guided by the contrasting nucleosome organization compared to other regions. By constructing an experimentally consistent near-atomic model of chromatin in the cellular environment, our approach provides a robust framework for future studies on nuclear compartmentalization, chromatin organization, and transcription regulation.

DOI： 10.1101/2024.10.22.619557

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Authorship：Lead author   Language：Others   Publishing type：Research paper (scientific journal)   Publisher：Society for Imaging Science & Technology  

Detection of Multiple Fluorescent Dyes Using Liquid Crystal Variable Retarder and Sparse Modeling

DOI： 10.2352/ei.2024.36.15.coimg-161

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Language：Others  

Responsible for pages：総ページ数：207, 担当ページ数：62-72   Language：English  

Event date： 2024.2

Language：Japanese  

Venue：東京   Country：Japan  

Event date： 2024.1

Language：English   Presentation type：Public lecture, seminar, tutorial, course, or other speech  

Venue：Kobe, RIKEN BDR   Country：Japan  

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Event date： 2023.11

Language：English  

Venue：名古屋   Country：Japan  

Higher-order genomic structures transformed during the transcription burst cycle

Event date： 2023.11

Language：English   Presentation type：Oral presentation (invited, special)  

Venue：Nagoya   Country：Japan  

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Other Link： https://www2.aeplan.co.jp/bsj2023/en-index.html

Event date： 2023.10

Language：English   Presentation type：Symposium, workshop panel (nominated)  

Country：Japan  

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Event date： 2023.10

Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Event date： 2023.9

Language：English   Presentation type：Symposium, workshop panel (nominated)  

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Event date： 2023.9

Language：English   Presentation type：Oral presentation (invited, special)  

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Event date： 2023.6

Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Event date： 2023.5

Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Event date： 2023.3

Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Event date： 2022.12

Language：Japanese  

Country：Other  

Event date： 2022.12

Language：Japanese   Presentation type：Oral presentation (invited, special)  

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Event date： 2021.2

Presentation type：Oral presentation (invited, special)  

Venue：Online  

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Language：English  

Venue：RIKEN IMS   Country：Japan  

Unveiling transcriptional dynamics in pluripotent stem cells through imaging analysis

Language：English  

Venue：Tokushima, Japan   Country：Other  

TALEN-mediated single-base-pair editing reveals the functional significance of an intergenic single nucleotide variant

Language：English  

Venue：Waltham, MA(USA)   Country：Other  

TALEN-mediated single-base-pair editing in PCS syndrome

Language：English  

Venue：CSHL, NY   Country：Other  

Stochastic promoter activation affects gene expression variability in murine embryonic stem cells

Language：Japanese  

Country：Other  

Language：English  

Country：Other  

Simultaneous live-imaging of gene position and its transcriptional activity in mouse embryonic stem cells

Language：Japanese  

Venue：熊本大学   Country：Other  

Language：English  

Venue：Awaji Yumebutai International Conference Center Awaji, Japan   Country：Other  

Simultaneous Live Imaging of the Transcription and Nuclear Position of Specific Genes

Language：Japanese  

Venue：広島大学   Country：Other  

Language：English  

Venue：Janelia Research Campus (VA), USA   Country：Other  

Simultaneous live imaging of the transcription and nuclear position of specific genes

Language：Japanese  

Country：Other  

Language：English  

Country：Other  

Simultaneous live imaging of a specific gene's transcription and dynamics.

Language：English  

Country：Other  

Active nuclear dynamics promotes heterochromatin clustering

Language：English  

Country：Other  

Relationship between nuclear dynamics and gene expression noise in pluripotent stem cell

Language：English  

Country：Other  

A challenging interdisciplinary approach to elucidate a mystery of remodeling process in nuclear architecture (Part II: Experiment)

Language：English  

Country：Other  

Single Base Pair Genome-Editing Technology and Its Future Applications

Language：English  

Venue：横浜   Country：Other  

Dynamic nuclear deformation induces nuclear architecture remodeling

Language：Japanese  

Country：Other  

Language：Japanese  

Country：Other  

Language：Japanese  

Country：Other  

Language：English  

Venue：熊本   Country：Other  

Relationship between kinetics of higher-order genomic structure and transcriptional activity

Language：Japanese  

Venue：神戸   Country：Other  

Relationship between kinetics of higher genome structure and transcription activity

Language：Japanese  

Venue：かでる2・7（北海道⽴道⺠活動センター）   Country：Other  

Comprehensive analysis of regulatory mechanisms of intrinsic gene expression noise in mouse embryonic stem cells

Language：Japanese  

Venue：大阪国際会議場   Country：Other  

Understanding Higher-order genomic structure using genome editing technology

Language：Japanese  

Country：Other  

Comprehensive analysis of regulatory mechanisms of gene expression noise in mouse embryonic stem cells

Language：Japanese  

Country：Other  

Development of comprehensive analysis technology for intercellular phenotypic heterogeneity

Language：Japanese  

Country：Other  

Role of dynamic nuclear deformation on genomic architecture reorganization

Language：Japanese  

Country：Other  

Language：English  

Venue：東京   Country：Other  

CRISPR library screening enables identification of transcriptional bursting related genes

Language：English  

Venue：Melbourne   Country：Other  

Transcriptional bursting induces gene expression heterogeneity in mouse embryonic stem cells

Language：English  

Venue：Basel   Country：Other  

Transcriptional bursting induces gene expression heterogeneity in mES cells

Language：Others  

Country：Other  

Language：Others  

Venue：オンライン   Country：Other  

Regulation of transcriptional bursting by chromatin potential

Language：Others  

Venue：オンライン   Country：Other  

Establishment of primitive endoderm differentiation reporter mouse ES cell line using CRISPR-Cas9

Language：Others  

Venue：Online   Country：Other  

Single-gene imaging reveals the relationship between the transcriptional activity of single genes and the formation of transcriptional regulatory factor clusters

Language：Others  

Venue：オンライン   Country：Other  

Relationship between higher-order genome structural dynamics and transcriptional dynamics

Language：Others  

Venue：オンライン   Country：Other  

Elucidation of the regulatory mechanism of transcriptional bursting that induces intercellular heterogeneity in gene expression

Language：Others  

Venue：オンライン   Country：Other  

Genome-wide kinetic properties of transcriptional bursting revealed by single cell analysis

Language：Others  

Venue：Online   Country：Other  

STREAMING-tag system: A novel technology to analyze the spatiotemporalrelationship between transcriptional regulators and transcriptionaldynamics at the single gene level

Language：Others  

Venue：郡山   Country：Other  

Relationship between Transcriptional Dynamics and Clusters of Transcriptional Regulators Revealed by Super-Resolution Imaging

Language：Others  

Venue：三島   Country：Other  

Language：Others  

Venue：Online   Country：Other  

Higher-order genomic structure and transcriptional dynamics

Language：Others  

Venue：国立遺伝学研究所   Country：Other  

Language：Others  

Venue：Sydney, Australia   Country：Other  

Revealing regulatory mechanisms of transcriptional dynamics by single-gene imaging system

Language：Others  

Venue：広島大学東広島キャンパス 学士会館   Country：Other  

Language：Others  

Venue：幕張メッセ、千葉   Country：Other  

Transcriptional activity-dependent formation of transcriptional regulatory factor condensate revealed by single gene imaging

Language：Others  

Venue：Ito Hall, The University of Tokyo   Country：Other  

Transcriptional dynamics regulated by non-genomic codes

Language：Others  

Venue：Zoom online   Country：Other  

Localization of transcriptional regulator condensates in transcriptional bursting revealed by STREAMING-tag system

Language：English  

Venue：オンライン   Country：Other  

Changes in higher-order genomeic structures during transcriptional dynamics revealed by multimodal chromatin tracing

Venue：Online  

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Venue：国立遺伝学研究所  

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Venue：広島大学東広島キャンパス 学士会館  

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Venue：Koriyama, Fukushima  

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Venue：Makuhari Messe, Chiba  

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Venue：三島  

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Language：English   Presentation type：Public lecture, seminar, tutorial, course, or other speech  

Venue：RIKEN IMS   Country：Japan  

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Venue：Ito Hall, The University of Tokyo  

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Language：English   Presentation type：Oral presentation (invited, special)  

Venue：Tsukuba   Country：Japan  

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Venue：Online  

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Venue：Sydney, Australia  

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Language：English   Presentation type：Symposium, workshop panel (nominated)  

Venue：online  

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Venue：Zoom online  

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Language：Japanese  

CRISPR-Cas9システムを利用したゲノム編集技術は、基礎研究利用に留まらず、医療応用や、有用畜産物の作出など、広く革新をもたらしている。一方で、DNAを切断しないCas9変異体を用いることで、ゲノム編集とは異なる応用も可能である。その1つが、CRISPR-Cas9システムを利用した特定遺伝子座のライブイメージングである。本稿では、特定ゲノム領域の可視化の意義、CRISPR-Cas9による生細胞イメージングのツールボックス、さらに本技術を利用した応用研究を紹介し、今後の展望も踏まえて議論する。(著者抄録)

Language：Japanese  

Comprehensive Analysis of Transcriptional Bursting in Mammalian Cells

DOI： 10.2142/biophys.61.171

Language：Japanese  

光学顕微鏡が普及し、肉眼では見えない細胞などの微細構造を観察できるようになり、多くの新しい生命現象の発見を支えてきた。とくに蛍光顕微鏡とその関連技術は、培養細胞のみならず、厚い組織切片、オルガノイド、生体などにおいて、特定分子の細胞内"動態"や発現量の"経時的変化"の測定を可能にし、創薬や病因同定などの医学分野にも深く関係している。本稿では、蛍光の原理および、1細胞レベルでのライブイメージングに用いられる蛍光顕微鏡、およびその派生技術である共焦点顕微鏡、多光子励起顕微鏡、ライトシート顕微鏡について解説する。さらに、1細胞レベルでの動態解析の重要性、蛍光ラベル化法、疾患オルガノイドを例にした医療への応用について解説する。(著者抄録)

Language：Japanese  

Functional analysis using CRISPR library

Language：Japanese  

DOI： 10.11477/mf.2425200621

Language：Japanese  

Identification of mutation causing cancer-prone hereditary disease using a novel method for the introduction of single-nucleotide substitution

Language：English  

Transcription activatorlike effector nucleases (TALENs) have recently arisen as effective tools for targeted genome engineering. Here, we report streamlined methods for the construction and evaluation of TALENs based on the Golden Gate TALEN and TAL Effector Kit' (Addgene). We diminished array vector requirements and increased assembly rates using six-module concatemerization. We altered the architecture of the native TALEN protein to increase nuclease activity and replaced the final destination vector with a mammalian expression/in vitro transcription vector bearing both CMV and T7 promoters. Using our methods, the whole process, from initiating construction to completing evaluation directly in mammalian cells, requires only 1week. Furthermore, TALENs constructed in this manner may be directly applied to transfection of cultured cells or mRNA synthesis for use in animals and embryos. In this article, we show genomic modification of HEK293T cells, human induced pluripotent stem cells, Drosophila melanogaster, Danio rerio and Xenopus laevis, using custom-made TALENs constructed and evaluated with our protocol. Our methods are more time efficient compared with conventional yeast-based evaluation methods and provide a more accessible and effective protocol for the application of TALENs in various model organisms.

DOI： 10.1111/gtc.12037

Language：Japanese  

Quantitative Imaging of Gene Expression Using Genome Editing Technology

Language：English  

Genome editing in cultured cells using TALEN

Language：Japanese  

Language：Japanese  

Language：Japanese  

Language：English  

Targetable nucleases, including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas), induce DNA double- strand breaks (DSBs) into user-defined sites. DSBs are immediately repaired through the evolutionarily conserved pathways of error-prone non-homologous end joining (NHEJ) or homology-directed repair (HDR). With the utilization of these repair processes, researchers have been able to disrupt specific genes, add exogenous DNA elements into intended genomic sites, introduce single-nucleotide substitutions, and perform many other applications. Consequently, this “genome editing” technology has revolutionized the life science field. In addition, this technology has the potential to improve agricultural products and be applicable to therapeutic use. Here, we will introduce a brief history of targetable nuclease-mediated genome editing and the applications of the tools that the technology provides. In this chapter, we will primarily focus on ZFNs and TALENs, which are artificial proteins composed of a specific DNA-binding domain and a restriction enzyme FokI DNAcleavage domain. We will also review the properties and construction methods of these nucleases.

DOI： 10.1007/978-4-431-55227-7_1

Type：Competition, symposium, etc. 

Type：Academic society, research group, etc. 

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Type：Academic society, research group, etc. 

Type：Competition, symposium, etc. 

Type：Scientific advice/Review 

Type：Competition, symposium, etc. 

Type：Competition, symposium, etc. 

Type：Competition, symposium, etc. 

Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Visiting lecture

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Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Visiting lecture

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Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Internet

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Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

Type：Newspaper, magazine

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Audience：General,　Scientific,　Company,　Civic organization,　Governmental agency

広島大が標的遺伝子の細胞内位置と活性を同時に可視化するROLEX技術、CRISPR/dCas9を活用

Simultaneous live imaging of a specific gene's transcription and dynamics

Simultaneous live imaging of a specific gene's transcription and dynamics

マウス胚性幹細胞における確率的なプロモーター活性化がNanog発現のばらつきに影響を与える

【山本研ゲノム編集アップデイト(3)】Wageningen大、DNAガイドを介した新規DNA切断現象を原核生物で発見

広島大と理研が人工ヌクレアーゼZFNで遺伝子挿入、ウニ初期胚1細胞の遺伝子発現を可視化