Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells
作者： Kirk R. Thomas , Mario R. Capecchi
DOI: 10.1016/0092-8674(87)90646-5
关键词: Transfection 、 Site-specific recombinase technology 、 Homology (biology) 、 Exon 、 Gene 、 Biology 、 Homologous recombination 、 Molecular biology 、 Gene targeting 、 Site-directed mutagenesis 、 General Biochemistry, Genetics and Molecular Biology
摘要: We mutated, by gene targeting, the endogenous hypoxanthine phosphoribosyl transferase (HPRT) in mouse embryo-derived stem (ES) cells. A specialized construct of neomycin resistance (neor) was introduced into an exon a cloned fragment Hprt and used to transfect ES Among G418r colonies, 1/1000 were also resistant base analog 6-thioguanine (6-TG). The G418r, 6-TGr cells all shown be Hprt- as result homologous recombination with exogenous, neor-containing, sequences. have compared gene-targeting efficiencies two classes neor-Hprt recombinant vectors: those that replace sequence exogenous insert sequence. targeting both vectors are strongly dependent upon extent homology between protocol described herein should useful for mutations any gene.
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