B-cells are isolated from mouse spleen. As upto my knowledge, mature B-cells won't proliferate in culture conditions. So is it possible to insert a gene (any gene, for a test condition) to such an isolated B-cells?
Dear S. Krishna,
There several methods to stably insert an exogenous gene into non-proliferating cells. Indeed even transfecction should give you stable random integration, but with a very reduced efficiency. I would try cloning your gene of interest in a lentiviral vector under a promoter already tested in your celltype. There're many references in the literature, why not read the following one to strart with:
"Molecular evidence of inefficient transduction of proliferating human B lymphocytes by VSV-pseudotyped HIV-1-derived lentivectors" Serafini M, Naldini L, Introna M. Virology. 2004 Aug 1;325(2):413-24.
BW
Dear S. Krishna,
There several methods to stably insert an exogenous gene into non-proliferating cells. Indeed even transfecction should give you stable random integration, but with a very reduced efficiency. I would try cloning your gene of interest in a lentiviral vector under a promoter already tested in your celltype. There're many references in the literature, why not read the following one to strart with:
"Molecular evidence of inefficient transduction of proliferating human B lymphocytes by VSV-pseudotyped HIV-1-derived lentivectors" Serafini M, Naldini L, Introna M. Virology. 2004 Aug 1;325(2):413-24.
BW
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