METHOD FOR CONSTRUCTING 293T CELL STRAIN WITH SITE-DIRECTED INSERTION OF BTDC GENE AND USE THEREOF

摘要

Disclosed are a method for constructing a 293T cell strain with site-directed insertion of the BTDC gene and a use thereof. The cell strain is obtained by constructing an sgRNA expression vector based on a CRISPR/Cas9 system according to the nucleotide sequence of an insertion site; constructing, according to an sgRNA action site, an expression cassette that contains the BTDC gene and can be integrated into a host genome; then co-transfecting the constructed sgRNA expression vector and a linearized expression cassette into 293T cells; and screening to obtain the 293T cell strain with site-directed insertion of the BTDC gene. The obtained cell strain can maintain the efficient and stable expression of the BTDC protein for a long period of time.