Genomic targeting with purified Cre recombinase.
Purified Cre recombinase protein introduced directly into cultured mammalian cells by lipofection catalyzes both site-specific chromosomal integration of a co-transfected lox targeting vector and precise excision of genomic DNA flanked by directly repeated lox sites. This procedure eliminates the need to transfect cre expression plasmids to activate recombination at lox sites. We used this simplified procedure to investigate the effect on targeting efficiency of both lox vector design and chromosomal position of the lox target. We show that such chromosomal position effects can exert at least a 50-fold per lox target difference in targeting efficiency in a human osteosarcoma cell line.
Baubonis
W
W
Biotechnology R & D, DuPont Merck Pharmaceutical Co., Wilmington, DE 19880-0400.
Sauer
B
B
D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004274:DNA, Recombinant / Q000378:metabolism*; D015894:Genome, Human; D006801:Humans; D019426:Integrases; D014162:Transfection; D014407:Tumor Cells, Cultured; D014764:Viral Proteins
D004274:DNA, Recombinant; D014764:Viral Proteins; C045073:Cre recombinase; D004254:DNA Nucleotidyltransferases; D019426:Integrases