Title:
PRODUCTION CELL AND PACKAGING CELL FOR RETROVIRAL VECTOR AND PREPARATION METHOD THEREFOR
Document Type and Number:
WIPO Patent Application WO/2021/218000
Kind Code:
A1
Abstract:
Provided are a production cell for producing a retroviral vector or a lentiviral vector carrying a nucleic acid fragment of interest and a preparation method therefor. The preparation method comprises using a sleeping beauty transposon system and a PiggyBac transposon system to integrate sequences of each gene into the genome of a host cell. Also provided are a nucleic acid fragment of interest and/or an envelope protein replacement system, a method, and a production cell for the retroviral vector. Also provided are a retroviral vector carrying the nucleic acid fragment of interest and use thereof in the preparation of a reagent for delivering the nucleic acid fragment of interest to a cell. Also provided are a packaging/production cell for preparing a lentiviral vector, a preparation method therefor and a method for preparing the lentiviral vector. The transcription of one or more of gag, pol and rev genes and coding sequences of VSV-G is under the sole control of a Tet-On inducible expression system or under the dual control of the Tet-On inducible expression system and a Cumate inducible expression system.
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Inventors:
XUE BOFU (CN)
YANG YINHUI (CN)
LIU KE (CN)
YANG YINHUI (CN)
LIU KE (CN)
Application Number:
PCT/CN2020/115522
Publication Date:
November 04, 2021
Filing Date:
September 16, 2020
Export Citation:
Assignee:
SHENZHEN EUREKA BIOTECHNOLOGY CO LTD (CN)
International Classes:
C12N15/86; C12N5/10; C12N7/00
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Other References:
GAO QIANG: "ESTABLISHMENT OF A PACKAGING CELL LINE FOR LENTIVIRUS VECTOR", CHINESE MASTER'S THESES FULL-TEXT DATABASE, TIANJIN POLYTECHNIC UNIVERSITY, CN, no. 1, 15 January 2007 (2007-01-15), CN , XP055862237, ISSN: 1674-0246
SCHUCHT, R. ET AL.: "A new generation of retroviral producer cells: predictable and stable virus production by Flp-mediated site-specific integration of retroviral vectors", MOL THER, vol. 14, no. 2, 2006, pages 285 - 292, XP005524740, DOI: 10.1016/j.ymthe.2005.12.003
LOEW, R. ET AL.: "A new PG13-based packaging cell line for stable production of clinical-grade self-inactivating gamma- retroviral vectors using targeted integration", GENE THER, vol. 17, no. 2, 2010, pages 272 - 280
YANT, S. R. ET AL.: "Mutational analysis of the N-terminal DNA-binding domain of sleeping beauty transposase: critical residues for DNA binding and hyperactivity in mammalian cells", MOL CELL BIOL, vol. 24, no. 20, 2004, pages 9239 - 9247, XP002466097, DOI: 10.1128/MCB.24.20.9239-9247.2004
LACOSTE, A. ET AL.: "An efficient and reversible transposable system for gene delivery and lineage-specific differentiation in human embryonic stem cells", CELL STEM CELL, vol. 5, no. 3, 2009, pages 332 - 342, XP055099142, DOI: 10.1016/j.stem.2009.07.011
YUSA, K. ET AL.: "A hyperactive piggyBac transposase for mammalian applications", PROC NATL ACAD SCI USA, vol. 108, no. 4, 2011, pages 1531 - 1536, XP055035951, DOI: 10.1073/pnas.1008322108
GOSSEN, M.H. BUJARD: "Tight control of gene expression in mammalian cells by tetracycline-responsive promoters", PROC NATL ACAD SCI U S A, vol. 89, no. 12, 1992, pages 5547 - 5551, XP002202393, DOI: 10.1073/pnas.89.12.5547
YU, H. ET AL.: "Inducible human immunodeficiency virus type 1 packaging cell lines", J VIROL, vol. 70, no. 7, 1996, pages 4530 - 4537
KAUL, M. ET AL.: "Regulated lentiviral packaging cell line devoid of most viral cis-acting sequences", VIROLOGY, vol. 249, no. 1, 1998, pages 167 - 174, XP002930834, DOI: 10.1006/viro.1998.9327
MULLICK, A. ET AL.: "The cumate gene-switch: a system for regulated expression in mammalian cells", BMC BIOTECHNOL, vol. 6, 2006, pages 43, XP021023579, DOI: 10.1186/1472-6750-6-43
AKEF, A. ET AL.: "Splicing promotes the nuclear export of beta-globin mRNA by overcoming nuclear retention elements", RNA, vol. 21, no. 11, 2015, pages 1908 - 1920
MICHAEL R. GREEN: "MolecjxLar Cloning: A Laboratory Manual", 2012, COLD SPRING HARBOR LABORATORY PRESS
SCHUCHT, R. ET AL.: "A new generation of retroviral producer cells: predictable and stable virus production by Flp-mediated site-specific integration of retroviral vectors", MOL THER, vol. 14, no. 2, 2006, pages 285 - 292, XP005524740, DOI: 10.1016/j.ymthe.2005.12.003
LOEW, R. ET AL.: "A new PG13-based packaging cell line for stable production of clinical-grade self-inactivating gamma- retroviral vectors using targeted integration", GENE THER, vol. 17, no. 2, 2010, pages 272 - 280
YANT, S. R. ET AL.: "Mutational analysis of the N-terminal DNA-binding domain of sleeping beauty transposase: critical residues for DNA binding and hyperactivity in mammalian cells", MOL CELL BIOL, vol. 24, no. 20, 2004, pages 9239 - 9247, XP002466097, DOI: 10.1128/MCB.24.20.9239-9247.2004
LACOSTE, A. ET AL.: "An efficient and reversible transposable system for gene delivery and lineage-specific differentiation in human embryonic stem cells", CELL STEM CELL, vol. 5, no. 3, 2009, pages 332 - 342, XP055099142, DOI: 10.1016/j.stem.2009.07.011
YUSA, K. ET AL.: "A hyperactive piggyBac transposase for mammalian applications", PROC NATL ACAD SCI USA, vol. 108, no. 4, 2011, pages 1531 - 1536, XP055035951, DOI: 10.1073/pnas.1008322108
GOSSEN, M.H. BUJARD: "Tight control of gene expression in mammalian cells by tetracycline-responsive promoters", PROC NATL ACAD SCI U S A, vol. 89, no. 12, 1992, pages 5547 - 5551, XP002202393, DOI: 10.1073/pnas.89.12.5547
YU, H. ET AL.: "Inducible human immunodeficiency virus type 1 packaging cell lines", J VIROL, vol. 70, no. 7, 1996, pages 4530 - 4537
KAUL, M. ET AL.: "Regulated lentiviral packaging cell line devoid of most viral cis-acting sequences", VIROLOGY, vol. 249, no. 1, 1998, pages 167 - 174, XP002930834, DOI: 10.1006/viro.1998.9327
MULLICK, A. ET AL.: "The cumate gene-switch: a system for regulated expression in mammalian cells", BMC BIOTECHNOL, vol. 6, 2006, pages 43, XP021023579, DOI: 10.1186/1472-6750-6-43
AKEF, A. ET AL.: "Splicing promotes the nuclear export of beta-globin mRNA by overcoming nuclear retention elements", RNA, vol. 21, no. 11, 2015, pages 1908 - 1920
MICHAEL R. GREEN: "MolecjxLar Cloning: A Laboratory Manual", 2012, COLD SPRING HARBOR LABORATORY PRESS
Attorney, Agent or Firm:
SHENZHEN ZHONGYI UNION INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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