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Detection of functional protein domains by unbiased genome-wide forward genetic screening

cam.issuedOnline2018-04-18
dc.contributor.authorHerzog, Mareike
dc.contributor.authorPuddu, Fabio
dc.contributor.authorCoates, Julia
dc.contributor.authorGeisler, Nicola
dc.contributor.authorForment, Josep
dc.contributor.authorJackson, Stephen
dc.contributor.orcidHerzog, Mareike [0000-0001-9747-2327]
dc.contributor.orcidPuddu, Fabio [0000-0002-2033-5209]
dc.contributor.orcidForment, Josep [0000-0002-7797-2583]
dc.contributor.orcidJackson, Stephen [0000-0001-9317-7937]
dc.date.accessioned2019-02-26T18:36:48Z
dc.date.available2019-02-26T18:36:48Z
dc.date.issued2018-04-18
dc.description.abstractGenetic and chemo-genetic interactions have played key roles in elucidating the molecular mechanisms by which certain chemicals perturb cellular functions. Many studies have employed gene knockout collections or gene disruption/depletion strategies to identify routes for evolving resistance to chemical agents. By contrast, searching for point-mutational genetic suppressors that can identify separation- or gain-of-function mutations, has been limited even in simpler, genetically amenable organisms such as yeast, and has not until recently been possible in mammalian cell culture systems. Here, by demonstrating its utility in identifying suppressors of cellular sensitivity to the drugs camptothecin or olaparib, we describe an approach allowing systematic, large-scale detection of spontaneous or chemically-induced suppressor mutations in yeast and in haploid mouse embryonic stem cells in a short timeframe, and with potential applications in essentially any other haploid system. In addition to its utility for molecular biology research, this protocol can be used to identify drug targets and to predict mechanisms leading to drug resistance. Mapping suppressor mutations on the primary sequence or three-dimensional structures of protein suppressor hits provides insights into functionally relevant protein domains, advancing our molecular understanding of protein functions, and potentially helping to improve drug design and applicability.
dc.identifier.doi10.17863/CAM.37236
dc.identifier.issn2045-2322
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/290009
dc.language.isoeng
dc.publisherNature Publishing Group
dc.publisher.urlhttps://www.nature.com/articles/s41598-018-24400-4#article-info
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subject3101 Biochemistry and Cell Biology
dc.subject3102 Bioinformatics and Computational Biology
dc.subject3105 Genetics
dc.subject31 Biological Sciences
dc.subjectBiotechnology
dc.subjectHuman Genome
dc.subjectGenetics
dc.subjectGeneric health relevance
dc.titleDetection of functional protein domains by unbiased genome-wide forward genetic screening
dc.typeArticle
dcterms.dateAccepted2018-03-22
prism.number6161
prism.publicationDate2018
prism.publicationNameScientific Reports
prism.volume8
pubs.funder-project-idCancer Research UK (18796)
pubs.funder-project-idCancer Research UK (C6946/A24843)
pubs.funder-project-idWellcome Trust (203144/Z/16/Z)
pubs.funder-project-idWellcome Trust (206388/Z/17/Z)
rioxxterms.licenseref.startdate2018-04-18
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
rioxxterms.typeJournal Article/Review
rioxxterms.versionVoR
rioxxterms.versionofrecord10.1038/s41598-018-24400-4

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