| First Author | Wu S | Year | 2007 |
| Journal | Nat Genet | Volume | 39 |
| Issue | 7 | Pages | 922-30 |
| PubMed ID | 17572674 | Mgi Jnum | J:123007 |
| Mgi Id | MGI:3716237 | Doi | 10.1038/ng2060 |
| Citation | Wu S, et al. (2007) Toward simpler and faster genome-wide mutagenesis in mice. Nat Genet 39(7):922-30 |
| abstractText | 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 alpha, beta and gamma 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. |
