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Toward simpler and faster genome-wide mutagenesis in mice.

Nature genetics | Jul 28, 2007

http://www.ncbi.nlm.nih.gov/pubmed/17572674

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.

Pubmed ID: 17572674 RIS Download

Mesh terms: Animals | Cadherins | Female | Genome | Male | Mice | Mice, Inbred C57BL | Mice, Knockout | Molecular Sequence Data | Mutagenesis | Recombination, Genetic | Translocation, Genetic

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Mouse Genome Informatics (Data, Gene Annotation)

Expression Atlas of the Marmoset (Data, Gene Annotation)

Mouse Genome Informatics Transgenes (Data, Gene Annotation)

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