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Kawabe Yoshinori Last modified date：2021.11.02

Associate Professor / Molecular and Biochemical Systems Engineering
Department of Chemical Engineering
Faculty of Engineering


Graduate School
Department of Chemical Systems and Engineering
Graduate School of Engineering
Undergraduate School
Department of Materials Science and Engineering
School of Engineering


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Homepage
https://kyushu-u.pure.elsevier.com/en/persons/kawabe-yoshinori
　Reseacher Profiling Tool　Kyushu University Pure
http://www.chem-eng.kyushu-u.ac.jp/lab3/index.html
Phone
092-802-2753
Fax
092-802-2793
Academic Degree
Doctor of Engineering
Country of degree conferring institution (Overseas)
No
Field of Specialization
Biochemical Engineering
Total Priod of education and research career in the foreign country
00years02months
Research
Research Interests
  • Development and application of microalgae biotechnology
    keyword : Algae, Microalgae, Chlamydomonas reinhardtii, transgene integration, gene retargeting, recombinant protein production
    2017.04.
  • Mass production sysytem for the biopharmaceutical proteins by transgenic avian bioreactor
    keyword : transgenic avian bioreactor, biopharmaceuticals, recombinant proteins, monoclonal antibody, retroviral vector
    2004.04.
  • development of maintainance and efficient derivation for pluripotential stem cells
    keyword : pluripotential stem cells, genetically manipulated animals, feeder layer, tissue engineering
    2008.04.
Academic Activities
Books
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1. Kawabe Y, Kamihira M, Antibody Expression and Production - Chapter 6 Production of Antibody by Transgenic Avians　Cell Engineering Volume 7 SpringerLink, pp121-141, 2011.04.
Reports
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Papers
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1. Yoshinori Kawabe, Akitsu Hotta, Ken-ichiro Ono, Kazuhisa Esaka, Ken-ichi Nishijima, Masamichi Kamihira, and Shinji Iijima, Production of chimeric antibodies by transgenic chicken bioreactor, Animal Cell Technology: Basic & Applied Aspects, 14,307-314, 2006.05.
2. Kawabe Y, Kamihira M, Ono K, Kyogoku K, Nishijima K, Iijima S, Production of scFv-Fc Fusion Protein by Genetically Manipulated Quails, J Biosci Bioeng , doi:10.1263/jbb.102.297, 102, 4, 297-303, 2006.10.
3. Kawabe Y, Naka T, Ando-Noumi N, Matsumoto H, Ono K, Nishijima K, Kamihira M, Iijima S, Transport of human Immunoglobulin G and Fc-Fusion Proteins to Chicken Egg Yolk, J Biosci Bioen, doi:10.1263/jbb.102.518, 102, 6, 518-523, 2006.12.
4. Kameyama Y, Kawabe Y, Ito A, Kamihira M, Antibody-dependent gene transduction using gammaretroviral and lentiviral vectors pseudotyped with chimeric vesicular stomatitis virus glycoprotein, J Virol Methods, doi:10.1016/j.jviromet.2008.06.013, 153, 1, 49-54, 2008.06.
5. Kawabe Y, Naka T, Komatsu H, Nishijima K, Iijima S, Kamihira M., Retroviral gene transduction into chicken embryo gonads through blood circulation, J Biosci Bioeng, 106(6):598-601, 2008.12.
6. Kamihira M, Kawabe Y, Shindo T, Ono K, Esaka K, Yamashita T, Nishijima K, Iijima S, Production of chimeric monoclonal antibodies by genetically manipulated chickens, J Biotechnol. , 141(1-2):18-25., 2009.04.
7. Penno CA, Kawabe Y, Ito A, Kamihira M., Production of recombinant human erythropoietin/Fc fusion protein by genetically manipulated chickens, Transgenic Res. , 19(2):187-95., 2010.04.
8. Kameyama Y, Kawabe Y, Ito A, Kamihira M., An accumulative site-specific gene integration system using Cre recombinase-mediated cassette exchange., Biotechnol Bioeng. , 105(6):1106-14., 2010.04.
9. Huang S, Kawabe Y, Ito A, Kamihira M , Cre recombinase-mediated site-specific modification of a celluar genome using an integrase-defective retroviral vector, Biotechnol Bioeng, 107: 717-29, 2010.11.
10. Yamamoto H, Kawabe Y, Ito A, Kamihira M, Enhanced liver functions in mouse hepatoma cells by induced overexpression of liver-enriched transcription factors., Biochem Eng J, doi:10.1016/j.bej.2011.10.004, 60, 15, 67-73, 2012.01.
11. Huang S, Kawabe Y, Ito A, Kamihira M, Adeno-associated virus Rep-mediated targeting of integrase-defective retroviral vector DNA circles into human chromosome 19., Biochem Biophys Res Commun, doi:10.1016/j.bbrc.2011.11.059, 417, 1, 78-83, 2012.01.
12. Obayashi H, Kawabe Y, Makitsubo H, Watanabe R, Kameyama Y, Huang S, Takenouchi Y, Ito A, Kamihira M, Accumulative gene integration into a pre-determined site using Cre/loxP., J Biosci Bioeng, doi:10.1016/j.jbiosc.2011.10.027, 113, 3, 381-388, 2012.05.
13. Kawabe Yoshinori, Makitsubo H, Kameyama Y, Huang S, Akira Ito, Masamichi Kamihira, Repeated integration of antibody genes into a pre-selected chromosomal locus of CHO cells using an accumulative site-specific gene integration system., Cytotechnology, DOI: 10.1007/s10616-011-9397-y, 64(3)、267-279, 2012.05.
14. Kawabe Y, Hayashida Y, Numata K, Harada S, Hayashida Y, Ito A, Kamihira M , Oral immunotherapy for pollen allergy using T-cell epitope-containing egg white derived from genetically manipulated chickens, PLOS ONE, doi:10.1371/journal.pone.0048512, 7, e48512, 2012.10.
15. Yoshinori Kawabe, Takanori Inao, Shodai Komatsu, Akira Ito, Masamichi Kamihira, Cre-mediated cellular modification for establishing producer CHO cells of recombinant scFv-Fc, BMC Proceedings, 10.1186/1753-6561-9-S9-P5, 9(Suppl 9):P5 , 2015.12.
16. Yoshinori Kawabe, Takuya Shimomura, Shuohao Huang, Suguru Imanishi, Akira Ito, Masamichi Kamihira, Targeted transgene insertion into the CHO cell genome using Cre recombinase-incorporating integrase-defective retroviral vectors., Biotechnol Bioeng., 10.1002/bit.25923, 113, 7, 1600-1610, 2016.01.
17. Yoshinori Kawabe, Takanori Inao, Shodai Komatsu, Guan Huang, Akira Ito, Takeshi Omasa, Masamichi Kamihira, Improved recombinant antibody production by CHO cells using a production enhancer DNA element with repeated transgene integration at a predetermined chromosomal site., J Biosci Bioeng., 10.1016/j.jbiosc.2016.10.011, 123, 3, 390-397, 2016.11.
18. Kawabe Yoshinori, Takanori Inao, Shodai Komatsu, Guan Huang, Akira Ito, Takeshi Omasa, Masamichi Kamihira, Improved recombinant antibody production by CHO cells using a production enhancer DNA element with repeated transgene integration at a predetermined chromosomal site, Journal of Bioscience and Bioengineering, 10.1016/j.jbiosc.2016.10.011, 123, 3, 390-397, 2017.03, Chinese hamster ovary (CHO) cells are one of the most useful host cell lines for the production of biopharmaceutical proteins. Although a series of production processes have been refined to improve protein productivity and cost performance, establishing producer cells is still time-consuming and labor-intensive. Recombinase-mediated site-specific gene integration into a predetermined chromosomal locus may enable predictable protein expression, reducing the laborious process of cell screening. We previously developed an accumulative site-specific gene integration system (AGIS) using Cre recombinase and mutated loxP sites for transgene integration and amplification in the CHO cell genome. Epigenetic modifier elements such as insulators are effective DNA cis-regulatory elements for stabilizing transgene expression. Here, we attempted to enhance transgene expression in recombinant CHO cells generated by AGIS using a production enhancer DNA element (PE) derived from the CHO genome. The PE was introduced into an expression unit for a recombinant scFv-Fc antibody. The effect on scFv-Fc productivity of PE position and orientation within the transgene was evaluated, while keeping the background chromosomal structure constant. For the optimal PE arrangement, scFv-Fc productivity was enhanced 2.6-fold compared with an expression unit without a PE. The enhancing effect of the PE on transgene expression was also observed when two or three PE-flanked expression units were inserted as tandem repeats. These results indicate that AGIS using the PE-flanked expression unit is a promising approach for establishing producer cell lines for biopharmaceutical protein production..
19. Xue Wang, Kawabe Yoshinori, Risa Kato, Takeshi Hada, Akira Ito, Yoshimasa Yamana, Masako Kondo, Masamichi Kamihira, Accumulative scFv-Fc antibody gene integration into the hprt chromosomal locus of Chinese hamster ovary cells, Journal of Bioscience and Bioengineering, 10.1016/j.jbiosc.2017.05.017, 124, 5, 583-590, 2017.11, We have previously developed an accumulative site-specific gene integration system (AGIS) using Cre-recombinase and mutated loxP sites. AGIS enables repeated transgene integration into a predetermined chromosomal site in mammalian cells. However, the process of establishing cells with multiple integrated copies of the transgene is still time-consuming. In the present study, we describe an improved version of AGIS that facilitates and accelerates the establishment of high-producer Chinese hamster ovary (CHO) cells. Two donor vectors were simultaneously introduced into the cells in a single transfection. Cells with successfully targeted transgene integration were screened based on a change in the color of the reporter fluorescent protein that they express. Repeated rounds of integration allowed the transgene copy number to be increased. As a model, an scFv-Fc antibody gene was integrated into the hprt locus of the CHO cell genome. After three rounds of integration, a high-producer CHO cell clone with six copies of the scFv-Fc gene was successfully established. scFv-Fc productivity was approximately four-fold greater than a control cell line harboring a single copy of the transgene. This newly designed AGIS procedure should facilitate the development of producer cells suitable for biopharmaceutical protein production..
20. Yoshinori Kawabe, Takuya Shimomura, Shuohao Huang, Suguru Imanishi, Akira Ito, Masamichi Kamihira, Development of retroviral vectors capable of site-specific gene insertion together with protein delivery, BMC Proceedings, https://doi.org/10.1186/s12919-018-0097-x, 2018, 12(Suppl 1):P-327, 2018.03.
21. Yoshinori Kawabe, Shinya Komatsu, Shodai Komatsu, Mai Murakami, Akira Ito, Tetsushi Sakuma, Takahiro Nakamura, Takashi Yamamoto, Masamichi Kamihira, Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems, J Biosci Bioeng, 10.1016/j.jbiosc.2017.12.003, 125, 5, 599-605, 2018.05.
22. Xue Wang, Yoshinori Kawabe, Takeshi Hada, Akira Ito, Masamichi Kamihira, Cre-mediated transgene integration in Chinese hamster ovary cells using minicircle DNA vectors, Biotechnology Journal, 10.1002/biot.201800063, 13, 7, e1800063, 2018.05.
23. Ming Shi, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira, Targeted knock-in into the OVA locus of chicken cells using CRISPR/Cas9 system with homology-independent targeted integration, Journal of Bioscience and Bioengineering, 10.1016/j.jbiosc.2019.09.011, 129, 3, 363-370, 2020.03, It is anticipated that transgenic avian species will be used as living bioreactors for the production of biopharmaceutical proteins. Precise tissue-specific expression of exogenous genes is a major challenge for the development of avian bioreactors. No robust vector is currently available for highly efficient and specific expression. In recent years, genome-editing techniques such as the CRISPR/Cas9 system have emerged as efficient and user-friendly genetic modification tools. Here, to apply the CRISPR/Cas9 system for the development of transgenic chickens, guide RNA sequences (gRNAs) of the CRISPR/Cas9 system for the ovalbumin (OVA) locus were evaluated for the oviduct-specific expression of exogenous genes. An EGFP gene expression cassette was introduced into the OVA locus of chicken DF-1 and embryonic fibroblasts using the CRISPR/Cas9 system mediated by homology-independent targeted integration. For the knock-in cells, EGFP expression was successfully induced by activation of the endogenous OVA promoter using the dCas9-VPR transactivation system. The combination of gRNAs designed around the OVA TATA box was important to induce endogenous OVA gene expression with high efficiency. These methods provide a useful tool for studies on the creation of transgenic chicken bioreactors and the activation of tissue-specific promoters..
24. Ryusei Iwao, Yoshinori Kawabe, Mai Murakami, Akira Ito, Masamichi Kamihira, Targeted Knock-in of Transgenes into the CHO Cell Genome Using CRISPR-mediated Integration Systems, MATEC Web of Conferences, https://doi.org/10.1051/matecconf/202133307001, 333, 07001, The 18th Asian Pacific Confederation of Chemical Engineering Congress (APCChE 2019), 2021.01.
25. Feiyang Zheng, Yoshinori Kawabe, Mai Murakami, Mamika Takahashi, Shoichiro Yoshida, Akira Ito, Masamichi Kamihira, Retrotransposon-mediated Gene Transfer for Animal Cells, MATEC Web of Conferences, https://doi.org/10.1051/matecconf/202133307002, 333, 07002, The 18th Asian Pacific Confederation of Chemical Engineering Congress (APCChE 2019), 2021.01.
26. Kazuki Shirakawa, Yoshinori Kawabe, Guan Huang, Akira Ito, Masamichi Kamihira, Targeted Gene Integration into Nuclear Genome of Microalgae Using Cre/loxP Recombination System, MATEC Web of Conferences, https://doi.org/10.1051/matecconf/202133307003, 333, 07003, The 18th Asian Pacific Confederation of Chemical Engineering Congress (APCChE 2019), 2021.01.
27. Feiyang Zheng, Yoshinori Kawabe, Mai Murakami, Mamika Takahashi, Kyoka Nishihata, Souichiro Yoshida, Akira Ito, Masamichi Kamihira, LINE-1 vectors mediate recombinant antibody gene transfer by retrotransposition in Chinese hamster ovary cells, Biotechnol J, https://doi.org/10.1002/biot.202000620, 16, 7, e2000620, 2021.07, [URL].
28. Guan Huang$, Yoshinori Kawabe$, Kazuki Shirakawa, Tatsuki Akiyama, Masamichi Kamihira ($Co-1st author), Novel transgenic Chlamydomonas reinhardtii strain with retargetable genomic transgene integration using Cre-loxP system  , J Biosci Bioeng, https://doi.org/10.1016/j.jbiosc.2021.07.006, in press, 2021.07.
29. Guan Huang, Yoshinori Kawabe, Kazuki Shirakawa, Tatsuki Akiyama, Masamichi Kamihira, Novel transgenic Chlamydomonas reinhardtii strain with retargetable genomic transgene integration using Cre-loxP system., Journal of bioscience and bioengineering, 10.1016/j.jbiosc.2021.07.006, 132, 5, 469-478, 2021.11, The use of Chlamydomonas for biofuel and biopharmaceutical production has been anticipated. However, the genetic engineering technology for Chlamydomonas is not as advanced as that for other organisms. Here, we established transgenic Chlamydomonas strains capable of high and stable transgene expression. The established cells exhibited stable reporter gene expression at a high level throughout long-term culture (∼60 days), even in the absence of drug pressure. The transgene insertion sites in the cell genome that may be suitable for exogenous gene expression were identified. Because the transgene contains a loxP site, the cells can be used as founders for retargeting other transgenes using the Cre-loxP system to generate transgenic Chlamydomonas producing useful substances. As a model biopharmaceutical gene, an interferon expression cassette was integrated into the genomic locus of the cells using Cre recombinase. The transgenic cells stably produced interferon protein in medium for 12 passages under non-selective conditions. These results indicate that the Chlamydomonas cells established in this study can serve as valuable and powerful tools not only for basic research on microalgae but also for the rapid establishment of cell lines expressing exogenous genes..
Presentations
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1. Huang Guan, Kawabe Yoshinori, Shirakawa Kazuki, Akiyama Tatsuki, Kamihira Masamichi, [PB212] Targeted integration of transgene into a pre-defined genomic locus of Chlamydomonas reinhardtii , 化学工学会第86年会, 2021.03.
2. Yoshinori Kawabe, Hirotaka Nishina, Ryusei Iwao, Masamichi Kamihira, Generation of a New CHK Cell Line with Inducible Differentiation for Enhanced Expression of Recombinant Antibody, JAACT2020 Fuchu, 2020.11.
3. Yoshinori Kawabe, Shinya Komatsu, Feiyang Zheng, Akira Ito, Masamichi Kamihira, Targeted knock-in into CHO cell genome using genome editing tools, 26th European Society for Animal Cell Technology (ESACT) Meeting, 2019.05.
4. Yoshinori Kawabe, Xue Wang, Takeshi Hada, Akio Kuno, Akira Ito, Masamichi Kamihira, Targeted scFv-Fc antibody gene integration into CHO cell genome using minicircle DNA vectors, JAACT2018 Tsukuba, 2018.11.
5. Yoshinori Kawabe, Takuya Shimomura, Shuohao Huang, Suguru Imanishi, Akira Ito, Masamichi Kamihira, Development of retroviral vectors capable of site-specific gene insertion together with protein delivery, 25th European Society for Animal Cell Technology (ESACT) Meeting , 2017.05.
6. Yoshinori Kawabe, Takanori Inao, Shodai Komatsu, Guan Huang, Akira Ito, Masamichi Kamihira, Improved scFv-Fc productivity in CHO cells with tandem-repeat expression units flanked by a production enhancer element, The 29th Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2016 Kobe), 2016.11.
7. Kawabe Yoshinori, Takanori Inao, Shodai Komatsu, Akira Ito, Masamichi Kamihira, Cre-mediated cellular modification for establishing producer CHO cells of recombinant scFv-Fc, ESACT ("European Society for Animal Cell Technology" Meeting) 2015, 2015.06.
8. Yoshinori Kawabe, Yuuki Hayashida, Kensaku Numata, Kenta Okuzono, Reina Obata, Akira Ito, Masamichi Kamihira, Egg-based oral immunotherapy with genetically manipulated chickens producing T-cell epitopes against Japanese cedar pollinosis, YABEC2014, 2014.11.
Membership in Academic Society
  • The Society for Biotechnology, Japan
  • JAPANESE ASSOCIATION FOR ANIMAL CELL TECHNOLOGY
  • THE MOLECULAR BIOLOGY SOCIETY OF JAPAN
  • THE SOCIETY OF CHEMICAL ENGINEERS, JAPAN
  • The Society of Biotechnology, Japan
  • The Society of Chemical Engineers, Japan
  • Japanese Association for Animal Cell Technology
  • The Molecular Biology Society of Japan
Educational
Other Educational Activities
  • 2020.11.
  • 2010.07.


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