A conditional knockout resource for the genome-wide study of mouse gene function

William C. Skarnes (), Barry Rosen, Anthony P. West, Manousos Koutsourakis, Wendy Bushell, Vivek Iyer, Alejandro O. Mujica, Mark Thomas, Jennifer Harrow, Tony Cox, David Jackson, Jessica Severin, Patrick Biggs, Jun Fu, Michael Nefedov, Pieter J. de Jong, A. Francis Stewart and Allan Bradley
Additional contact information
William C. Skarnes: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Barry Rosen: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Anthony P. West: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Manousos Koutsourakis: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Wendy Bushell: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Vivek Iyer: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Alejandro O. Mujica: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Mark Thomas: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Jennifer Harrow: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Tony Cox: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
David Jackson: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Jessica Severin: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Patrick Biggs: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton
Jun Fu: Biotechnologisches Zentrum, TU Dresden
Michael Nefedov: Children’s Hospital Oakland Research Institute
Pieter J. de Jong: Children’s Hospital Oakland Research Institute
A. Francis Stewart: Biotechnologisches Zentrum, TU Dresden
Allan Bradley: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton

Nature, 2011, vol. 474, issue 7351, 337-342

Abstract: 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.

Date: 2011
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DOI: 10.1038/nature10163

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