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- Title
A conditional knockout resource for the genome-wide study of mouse gene function.
- Authors
Skarnes, William C.; Rosen, Barry; West, Anthony P.; Koutsourakis, Manousos; Bushell, Wendy; Iyer, Vivek; Mujica, Alejandro O.; Thomas, Mark; Harrow, Jennifer; Cox, Tony; Jackson, David; Severin, Jessica; Biggs, Patrick; Fu, Jun; Nefedov, Michael; de Jong, Pieter J.; Stewart, A. Francis; Bradley, Allan
- Abstract
Gene targeting in embryonic stem cells has become the principal technology for manipulation of the mouse genome, offering unrivalled accuracy in allele design and access to conditional mutagenesis. To bring these advantages to the wider research community, large-scale mouse knockout programmes are producing a permanent resource of targeted mutations in all protein-coding genes. Here we report the establishment of a high-throughput gene-targeting pipeline for the generation of reporter-tagged, conditional alleles. Computational allele design, 96-well modular vector construction and high-efficiency gene-targeting strategies have been combined to mutate genes on an unprecedented scale. So far, more than 12,000 vectors and 9,000 conditional targeted alleles have been produced in highly germline-competent C57BL/6N embryonic stem cells. High-throughput genome engineering highlighted by this study is broadly applicable to rat and human stem cells and provides a foundation for future genome-wide efforts aimed at deciphering the function of all genes encoded by the mammalian genome.
- Subjects
EMBRYONIC stem cells; GENE targeting; MUTAGENESIS; LABORATORY mice; GENOMICS; GENETIC engineering; GENE frequency
- Publication
Nature, 2011, Vol 474, Issue 7351, p337
- ISSN
0028-0836
- Publication type
Article
- DOI
10.1038/nature10163