Technical Report abstract
Nature Genetics 39, 922 - 930 (2007)
Published online: 17 June 2007 | doi:10.1038/ng2060
Toward simpler and faster genome-wide mutagenesis in mice
Sen Wu1, Guoxin Ying2, Qiang Wu2 & Mario R Capecchi1,2
Abstract
Here we describe a practical Cre-loxP and piggyBac transposon–based mutagenesis strategy to systematically mutate coding sequences and/or the vast noncoding regions of the mouse genome for large-scale functional genomic analysis. To illustrate this approach, we first created loxP-containing loss-of-function alleles in the protocadherin 
, 
and 
gene clusters (Pcdha, Pcdhb and Pcdhg). Using these alleles, we show that, under proper guidance, Cre-loxP site-specific recombination can mediate efficient trans-allelic recombination in vivo, facilitating the generation of large germline deletions and duplications including deletions of Pcdha, and Pcdha to Pcdhb, simply by breeding (that is, at frequencies of 5.5%–21.6%). The same breeding method can also generate designed germline translocations between nonhomologous chromosomes at unexpected frequencies of greater than 1%. By incorporating a piggyBac transposon to insert and to distribute loxP sites randomly throughout the mouse genome, we present a simple but comprehensive method for generating genome-wide deletions and duplications, in addition to insertional loss-of-function and conditional rescue alleles, again simply by breeding.
- Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah 84112, USA.
- Department of Human Genetics, University of Utah, Salt Lake City, Utah 84112, USA.
Correspondence to: Mario R Capecchi1,2 e-mail: mario.capecchi@genetics.utah.edu
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