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Manipulating the Mouse Genome Using Recombineering,

Kajal Biswas and Shyam K Sharan

Genetically engineered mouse models are indispensable for understanding the biological function of genes, understanding the genetic basis of human diseases, and for preclinical testing of novel therapies. Generation of such mouse models has been possible because of our ability to manipulate the mouse genome. Recombineering is a highly efficient, recombination-based method of genetic engineering that has revolutionized our ability to generate mouse models. Since recombineering technology is not dependent on the availability of restriction enzyme recognition sites, it allows us to modify the genome with great precision. It requires homology arms as short as 40 bases for recombination, which makes it relatively easy to generate targeting constructs to insert, change, or delete either a single nucleotide or a DNA fragment several kb in size; insert selectable markers or reporter genes; or add epitope tags to any gene of interest. In this review, we focus on the development of recombineering technology and its application to the generation of genetically engineered mouse models. High-throughput generation of gene targeting vectors, used to construct knockout alleles in mouse embryonic stem cells, is now feasible because of this technology. The challenge now is to use these “designer” mice to develop novel therapies to prevent, cure, or effectively manage some the most debilitating human diseases.