A gene-driven ENU-based approach to generating an allelic series in any gene

Mohamed Mohideen Quwailid, Alison Hugill, Terence Neil Dear, Lucie Vizor, Sara Wells, Emma Horner, Shelly Fuller, Jessica Weedon, Hamish McMath, Paul Woodman, David Edwards, David Campbell, Susan Rodger, Joanne Carey, Ann Roberts, Pete Glenister, Zuzanna Lalanne, Nick Parkinson, Emma L. Coghill, Richard McKeone & 10 others Sam Cox, John Willan, Andy Greenfield, David Keays, Saffron Brady, Nigel Spurr, Ian Gray, Jackie Hunter, Steve D.M. Brown, Roger D. Cox

Research output: Contribution to journalArticle

133 Citations (Scopus)

Abstract

N-ethyl-N-nitrosourea (ENU) introduces mutations throughout the mouse genome at relatively high efficiency. Successful high-throughput phenotype screens have been reported and alternative screens using sequence-based approaches have been proposed. For the purpose of generating an allelic series in selected genes by a sequence-based approach, we have constructed an archive of over 4000 DNA samples from individual F1 ENU-mutagenized mice paralleled by frozen sperm samples. Together with our previously reported archive, the total size now exceeds 6000 individuals. A gene-based screen of 27.4 Mbp of DNA, carried out using denaturing high-performance liquid chromatography (DHPLC), found a mutation rate of 1 in 1.01 Mbp of which 1 in 1.82 Mbp were potentially functional. Screening of whole or selected regions of genes on subsets of the archive has allowed us to identify 15 new alleles from 9 genes out of 15 tested. This is a powerful adjunct to conventional mutagenesis strategies and has the advantage of generating a variety of alleles with potentially different phenotypic outcomes that facilitate the investigation of gene function. It is now available to academic collaborators as a community resource.

LanguageEnglish
Pages585-591
Number of pages7
JournalMammalian Genome
Volume15
Issue number8
DOIs
Publication statusPublished - 1 Aug 2004

ASJC Scopus subject areas

  • Genetics

Cite this

Quwailid, M. M., Hugill, A., Dear, T. N., Vizor, L., Wells, S., Horner, E., ... Cox, R. D. (2004). A gene-driven ENU-based approach to generating an allelic series in any gene. Mammalian Genome, 15(8), 585-591. https://doi.org/10.1007/s00335-004-2379-z