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

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.

journal_name

Nature

journal_title

Nature

authors

Skarnes WC,Rosen B,West AP,Koutsourakis M,Bushell W,Iyer V,Mujica AO,Thomas M,Harrow J,Cox T,Jackson D,Severin J,Biggs P,Fu J,Nefedov M,de Jong PJ,Stewart AF,Bradley A

doi

10.1038/nature10163

subject

Has Abstract

pub_date

2011-06-15 00:00:00

pages

337-42

issue

7351

eissn

0028-0836

issn

1476-4687

pii

nature10163

journal_volume

474

pub_type

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