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Article: Increased efficiency of oligonucleotide-mediated gene repair through slowing replication fork progression

TitleIncreased efficiency of oligonucleotide-mediated gene repair through slowing replication fork progression
Authors
KeywordsSingle-stranded Oligonucleotide
Targeted gene repair
Thymidine
Issue Date2005
PublisherNational Academy of Sciences. The Journal's web site is located at http://www.pnas.org
Citation
Proceedings Of The National Academy Of Sciences Of The United States Of America, 2005, v. 102 n. 7, p. 2508-2513 How to Cite?
AbstractTargeted gene modification mediated by single-stranded oligonucleotides (SSOs) holds great potential for widespread use in a number of biological and biomedical fields, including functional genomics and gene therapy. By using this approach, specific genetic changes have been created in a number of prokaryotic and eukaryotic systems. In mammalian cells, the precise mechanism of SSO-mediated chromosome alteration remains to be established, and there have been problems in obtaining reproducible targeting efficiencies. It has previously been suggested that the chromatin structure, which changes throughout the cell cycle, may be a key factor underlying these variations in efficiency. This hypothesis prompted us to systematically investigate SSO-mediated gene repair at various phases of the cell cycle in a mammalian cell line. We found that the efficiency of SSO-mediated gene repair was elevated by ≈10-fold in thymidine-treated S-phase cells. The increase in repair frequency correlated positively with the duration of SSO/thymidine coincubation with host cells after transfection. We supply evidence suggesting that these increased repair frequencies arise from a thymidine-induced slowdown of replication fork progression. Our studies provide fresh insight into the mechanism of SSO-mediated gene repair in mammalian cells and demonstrate how its efficiency may be reliably and substantially increased.
Persistent Identifierhttp://hdl.handle.net/10722/48989
ISSN
2015 Impact Factor: 9.423
2015 SCImago Journal Rankings: 6.883
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWu, XSen_HK
dc.contributor.authorXin, Len_HK
dc.contributor.authorYin, WXen_HK
dc.contributor.authorShang, XYen_HK
dc.contributor.authorLu, Len_HK
dc.contributor.authorWatt, RMen_HK
dc.contributor.authorCheah, KSEen_HK
dc.contributor.authorHuang, JDen_HK
dc.contributor.authorLiu, DPen_HK
dc.contributor.authorLiang, CCen_HK
dc.date.accessioned2008-06-12T06:31:29Z-
dc.date.available2008-06-12T06:31:29Z-
dc.date.issued2005en_HK
dc.identifier.citationProceedings Of The National Academy Of Sciences Of The United States Of America, 2005, v. 102 n. 7, p. 2508-2513en_HK
dc.identifier.issn0027-8424en_HK
dc.identifier.urihttp://hdl.handle.net/10722/48989-
dc.description.abstractTargeted gene modification mediated by single-stranded oligonucleotides (SSOs) holds great potential for widespread use in a number of biological and biomedical fields, including functional genomics and gene therapy. By using this approach, specific genetic changes have been created in a number of prokaryotic and eukaryotic systems. In mammalian cells, the precise mechanism of SSO-mediated chromosome alteration remains to be established, and there have been problems in obtaining reproducible targeting efficiencies. It has previously been suggested that the chromatin structure, which changes throughout the cell cycle, may be a key factor underlying these variations in efficiency. This hypothesis prompted us to systematically investigate SSO-mediated gene repair at various phases of the cell cycle in a mammalian cell line. We found that the efficiency of SSO-mediated gene repair was elevated by ≈10-fold in thymidine-treated S-phase cells. The increase in repair frequency correlated positively with the duration of SSO/thymidine coincubation with host cells after transfection. We supply evidence suggesting that these increased repair frequencies arise from a thymidine-induced slowdown of replication fork progression. Our studies provide fresh insight into the mechanism of SSO-mediated gene repair in mammalian cells and demonstrate how its efficiency may be reliably and substantially increased.en_HK
dc.format.extent386 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherNational Academy of Sciences. The Journal's web site is located at http://www.pnas.orgen_HK
dc.relation.ispartofProceedings of the National Academy of Sciences of the United States of Americaen_HK
dc.rightsNational Academy of Sciences Proceedings. Copyright © National Academy of Sciences.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subjectSingle-stranded Oligonucleotideen_HK
dc.subjectTargeted gene repairen_HK
dc.subjectThymidineen_HK
dc.titleIncreased efficiency of oligonucleotide-mediated gene repair through slowing replication fork progressionen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0027-8424&volume=102&issue=7&spage=2508&epage=2513&date=2005&atitle=Increased+efficiency+of+oligonucleotide-mediated+gene+repair+through+slowing+replication+fork+progressionen_HK
dc.identifier.emailWatt, RM:rmwatt@hku.hken_HK
dc.identifier.emailCheah, KSE:hrmbdkc@hku.hken_HK
dc.identifier.emailHuang, JD:jdhuang@hkucc.hku.hken_HK
dc.identifier.authorityWatt, RM=rp00043en_HK
dc.identifier.authorityCheah, KSE=rp00342en_HK
dc.identifier.authorityHuang, JD=rp00451en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1073/pnas.0406991102en_HK
dc.identifier.pmid15695590-
dc.identifier.pmcidPMC548982en_HK
dc.identifier.scopuseid_2-s2.0-20044370284en_HK
dc.identifier.hkuros97946-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-20044370284&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume102en_HK
dc.identifier.issue7en_HK
dc.identifier.spage2508en_HK
dc.identifier.epage2513en_HK
dc.identifier.isiWOS:000227073100048-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridWu, XS=37059359400en_HK
dc.identifier.scopusauthoridXin, L=36060607800en_HK
dc.identifier.scopusauthoridYin, WX=8988889100en_HK
dc.identifier.scopusauthoridShang, XY=27968135600en_HK
dc.identifier.scopusauthoridLu, L=24332526400en_HK
dc.identifier.scopusauthoridWatt, RM=7102907536en_HK
dc.identifier.scopusauthoridCheah, KSE=35387746200en_HK
dc.identifier.scopusauthoridHuang, JD=8108660600en_HK
dc.identifier.scopusauthoridLiu, DP=8047815300en_HK
dc.identifier.scopusauthoridLiang, CC=7403280685en_HK

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