Gene-targeting strategies.
Gene targeting in mouse ES cells is a powerful method for studying gene function in vivo. For the novice, this combination of molecular biology, specialized tissue-culture cell lines, and mouse reproductive biology can be daunting. We present a straightforward, one might say constrained, guide for novices of gene targeting to generate a null allele in large or small protein-coding genes. The method we outline has evolved from years of experience of training and advising beginners on this powerful technology. We believe that a good design for a gene-targeting strategy ultimately saves time, money, and research animal lives. Once you feel comfortable with a fundamental knockout, we suggest you consider the new and exciting gene-targeting variations that can be used to address important biological questions (2).
Cheah
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SS
Department of Molecular Genetics, University of Texas M. D. Anderson Cancer Center, Houston, USA.
Behringer
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RR
D000818:Animals; D018390:Gene Targeting / Q000379:methods*; D051379:Mice