FIELD: biotechnology.
SUBSTANCE: described is a lentiviral vector transfer plasmid comprising a nucleotide sequence encoding a vector cassette TL20c, a nucleotide sequence encoding a packing signal; a nucleotide sequence encoding a central polypurine pathway; nucleotide sequence encoding the responsive Rev element; and a nucleotide sequence, coding a self-inactivated long terminal repeat; and where plasmid includes a multiple cloning site comprising four restriction endonuclease sites, where four restriction nuclease sites are selected from a group consisting of BstBI, MluI, NotI, ClaI, ApaI, XhoI, XbaI, HpaI, NheI, PacI, NsiI, SphI, Sma/Xma, AccI, BamHI and SphI. Disclosed is a cathode matrix for use in producing and/or transfecting a packaging cell line containing DNA fragments obtained from a vector cassette, having nucleotide sequence SEQ ID NO: 2. What is described is a packing cell line transfected with a concatemeric matrix according to any of claims 11, 12, to produce a stable producer cell line, where the packaging cell line is GPRG. Also described is a stable cell line-producer selected from a transfected packaging cell line according to claim 13.
EFFECT: invention provides a method for producing a stable producer cell line; obtaining stable lines of producer cells, such as presented in accordance with this invention, increases reproducibility and simplicity of creation of lentiviral effluents with high titre, simultaneously reducing problems of biosafety, and change of expressed envelope proteins determines tropism of produced virus.
17 cl, 20 dwg, 5 tbl, 7 ex
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Authors
Dates
2020-08-06—Published
2016-05-12—Filed