FIELD: biotechnology.
SUBSTANCE: invention relates to biotechnology. Disclosed is a method for the T cell genetic modification including: the interaction of an ex vivo T cell with a replication-incompetent recombinant retrovirus to form a reaction mixture for transduction, with the replication-incompetent recombinant retrovirus comprising: a) one or more pseudotyped elements on the surface of a replication-incompetent recombinant retrovirus, with one or more pseudotyped elements facilitating the binding to and fusion with the T cell of a replication-incompetent recombinant retrovirus; and b) a polynucleotide containing one or more transcription units, with each of which being functionally linked to a promoter active in T cells, with one or more transcription units encoding the first constructed signal polypeptide including the first chimeric antigen receptor containing an antigenic target area; a transmembrane domain; and an intracellular activation domain, and c) an activation element on the surface of a replication-incompetent recombinant retrovirus, with said activation element being a membranous polypeptide capable of binding to CD3 on the surface of the resting T cell and of activating the resting T cell and is not encoded by a polynucleotide in a replication-incompetent recombinant retrovirus, with the T cell interacting ex vivo with a replication-incompetent recombinant retrovirus within an interval of 15 minutes to 14 hours. Furthermore, the method is carried out without pre-stimulation ex vivo with said interaction facilitating the membrane fusion of the T cell with a replication-incompetent recombinant retrovirus for the production of a genetically modified T cell. Along with disclosed is a recombinant retrovirus.
EFFECT: invention enables T cell genetic modification.
23 cl, 21 dwg, 3 tbl, 13 ex
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Authors
Dates
2021-09-13—Published
2017-03-19—Filed