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- Title
Toward simpler and faster genome-wide mutagenesis in mice.
- Authors
Sen Wu; Guoxin Ying; Qiang Wu; Capecchi, Mario R.
- 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 α, β and γ 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.
- Publication
Nature Genetics, 2007, Vol 39, Issue 7, p922
- ISSN
1061-4036
- Publication type
Academic Journal
- DOI
10.1038/ng2060