Using Recombineering Technology to Create Genetically Engineered Mouse Models
Our ability to manipulate the mouse genome has been the key to the generation of suitable mouse models of human diseases. This has been achieved by introducing foreign DNA either to ectopically express genes or to render endogenous genes nonfunctional by gene targeting technology. Genomic manipulation has been revolutionized by the availability of bacterial artificial chromosomes and the development of recombineering technology, a recombination-based method of genetic engineering. These have not only helped to simplify and reduce the time involved in genetic modification but have also led to the generation of mouse models that were not feasible earlier. In this chapter, we describe the recombineering technology and discuss its applications to generate knockout, knockin, or conditional gene targeting constructs. In addition, we describe how recombineering can be used to insert, delete, or create subtle alterations in the large DNA segments that can be used to generate transgenic mouse models.
KeywordsEmbryonic Stem Cell Selectable Marker Internal Ribosome Entry Site Mixed Lineage Leukemia loxP Site
The authors thank Drs. Kajal Biswas, Suhwan Chang, Lino Tessarollo, and Sergey Kuznetsov for critical review of the manuscript and Tammy Schroyer of the Publication Department for the illustrations. This research was supported by the Center for Cancer Research, National Cancer Institute, Department of Human and Health Services.
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