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

Mohamed Mohideen Quwailid; Alison Hugill; 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; Sam Cox; John Willan; Andy Greenfield; David Keays; Saffron Brady; Nigel Spurr; Ian Gray; Jackie Hunter; Steve D.M. Brown; Roger D. Cox

doi:10.1007/s00335-004-2379-z

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

Mohamed Mohideen Quwailid, Alison Hugill, 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 McKeoneSam 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 journal › Article › peer-review

137 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.

Original language	English
Pages (from-to)	585-591
Number of pages	7
Journal	Mammalian Genome
Volume	15
Issue number	8
DOIs	https://doi.org/10.1007/s00335-004-2379-z
Publication status	Published or Issued - Aug 2004
Externally published	Yes

ASJC Scopus subject areas

Genetics

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Quwailid, M. M., Hugill, A., Dear, N., Vizor, L., Wells, S., Horner, E., Fuller, S., Weedon, J., McMath, H., Woodman, P., Edwards, D., Campbell, D., Rodger, S., Carey, J., Roberts, A., Glenister, P., Lalanne, Z., Parkinson, N., Coghill, E. L., ... 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

@article{89919887e4b0476889466dd101fd9a10,

title = "A gene-driven ENU-based approach to generating an allelic series in any gene",

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.",

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

year = "2004",

month = aug,

doi = "10.1007/s00335-004-2379-z",

language = "English",

volume = "15",

pages = "585--591",

journal = "Mammalian Genome",

issn = "0938-8990",

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Quwailid, MM, Hugill, A, Dear, N, Vizor, L, Wells, S, Horner, E, Fuller, S, Weedon, J, McMath, H, Woodman, P, Edwards, D, Campbell, D, Rodger, S, Carey, J, Roberts, A, Glenister, P, Lalanne, Z, Parkinson, N, Coghill, EL, McKeone, R, Cox, S, Willan, J, Greenfield, A, Keays, D, Brady, S, Spurr, N, Gray, I, Hunter, J, Brown, SDM & Cox, RD 2004, 'A gene-driven ENU-based approach to generating an allelic series in any gene', Mammalian Genome, vol. 15, no. 8, pp. 585-591. https://doi.org/10.1007/s00335-004-2379-z

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T1 - A gene-driven ENU-based approach to generating an allelic series in any gene

AU - Quwailid, Mohamed Mohideen

AU - Hugill, Alison

AU - Dear, Neil

AU - Vizor, Lucie

AU - Wells, Sara

AU - Horner, Emma

AU - Fuller, Shelly

AU - Weedon, Jessica

AU - McMath, Hamish

AU - Woodman, Paul

AU - Edwards, David

AU - Campbell, David

AU - Rodger, Susan

AU - Carey, Joanne

AU - Roberts, Ann

AU - Glenister, Pete

AU - Lalanne, Zuzanna

AU - Parkinson, Nick

AU - Coghill, Emma L.

AU - McKeone, Richard

AU - Cox, Sam

AU - Willan, John

AU - Greenfield, Andy

AU - Keays, David

AU - Brady, Saffron

AU - Spurr, Nigel

AU - Gray, Ian

AU - Hunter, Jackie

AU - Brown, Steve D.M.

AU - Cox, Roger D.

PY - 2004/8

Y1 - 2004/8

N2 - 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.

AB - 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.

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DO - 10.1007/s00335-004-2379-z

M3 - Article

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SN - 0938-8990

VL - 15

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EP - 591

JO - Mammalian Genome

JF - Mammalian Genome

IS - 8

ER -

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

Abstract

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