Animal models for disease: knockout, knock-in, and conditional mutant mice

Diseases with a genetic basis can be modeled with knockout, knock-in, and conditional mutant gene-targeted mice. In the following, we provide detailed protocols for gene targeting. Gene targeting of embryonic stem cells can be accomplished by laboratories equipped for tissue culture. Alternatively, many gene-targeting services divide the work of targeting with a customer lab. In this collaborative situation, knowledge of the entire process helps ensure a successful outcome. The construction of chimeras for germ-line transmission is not described here, because this procedure is beyond the means of most laboratories, typically is provided by transgenic core facilities, and is best learned through hands-on demonstration.