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Article: Active transcription promotes single-stranded oligonucleotide mediated gene repair

TitleActive transcription promotes single-stranded oligonucleotide mediated gene repair
Authors
Keywordsλ-Red
Gene therapy
Recombination
Strand bias
Issue Date2007
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
Citation
Biochemical And Biophysical Research Communications, 2007, v. 353 n. 1, p. 33-39 How to Cite?
AbstractThe λ-Red-mediated recombination has been exploited as an efficient means for DNA manipulation. We previously reported that replication plays a pivotal role during this process. Replication direction dictates strand bias, such that single stranded oligonucleotide (SSO) with sequence corresponding to the nascent lagging strand directs higher levels of recombinant formation compared to its complementary SSO. In addition, the Escherichia coli methyl-directed mismatch repair system impedes efficient SSO-mediated site-specific gene repair. However, the role of transcription in determining strand bias and recombination efficiency is unclear. To address the potential role of transcriptional processes, we constructed plasmid substrates that harbor a mutant antibiotic reporter under the control of an inducible promoter. We found that transcription activation can promote recombinant formation to more than 10-folds whilst it has negligible effect on strand bias. Our findings provide evidence for a role of transcription in SSO-mediated gene repair process. © 2006 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/91042
ISSN
2023 Impact Factor: 2.5
2023 SCImago Journal Rankings: 0.770
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorHuen, MSYen_HK
dc.contributor.authorLu, LYen_HK
dc.contributor.authorLiu, DPen_HK
dc.contributor.authorHuang, JDen_HK
dc.date.accessioned2010-09-17T10:12:10Z-
dc.date.available2010-09-17T10:12:10Z-
dc.date.issued2007en_HK
dc.identifier.citationBiochemical And Biophysical Research Communications, 2007, v. 353 n. 1, p. 33-39en_HK
dc.identifier.issn0006-291Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/91042-
dc.description.abstractThe λ-Red-mediated recombination has been exploited as an efficient means for DNA manipulation. We previously reported that replication plays a pivotal role during this process. Replication direction dictates strand bias, such that single stranded oligonucleotide (SSO) with sequence corresponding to the nascent lagging strand directs higher levels of recombinant formation compared to its complementary SSO. In addition, the Escherichia coli methyl-directed mismatch repair system impedes efficient SSO-mediated site-specific gene repair. However, the role of transcription in determining strand bias and recombination efficiency is unclear. To address the potential role of transcriptional processes, we constructed plasmid substrates that harbor a mutant antibiotic reporter under the control of an inducible promoter. We found that transcription activation can promote recombinant formation to more than 10-folds whilst it has negligible effect on strand bias. Our findings provide evidence for a role of transcription in SSO-mediated gene repair process. © 2006 Elsevier Inc. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/descriptionen_HK
dc.relation.ispartofBiochemical and Biophysical Research Communicationsen_HK
dc.subjectλ-Reden_HK
dc.subjectGene therapyen_HK
dc.subjectRecombinationen_HK
dc.subjectStrand biasen_HK
dc.subject.meshDNA Damage - geneticsen_HK
dc.subject.meshDNA Repair - geneticsen_HK
dc.subject.meshDNA, Bacterial - geneticsen_HK
dc.subject.meshEscherichia coli - geneticsen_HK
dc.subject.meshRecombination, Genetic - geneticsen_HK
dc.subject.meshTargeted Gene Repair - methodsen_HK
dc.subject.meshTranscriptional Activation - geneticsen_HK
dc.titleActive transcription promotes single-stranded oligonucleotide mediated gene repairen_HK
dc.typeArticleen_HK
dc.identifier.emailHuen, MSY:huen.michael@hku.hken_HK
dc.identifier.emailHuang, JD:jdhuang@hkucc.hku.hken_HK
dc.identifier.authorityHuen, MSY=rp01336en_HK
dc.identifier.authorityHuang, JD=rp00451en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.bbrc.2006.11.146en_HK
dc.identifier.pmid17174937-
dc.identifier.scopuseid_2-s2.0-33845608839en_HK
dc.identifier.hkuros128577-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33845608839&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume353en_HK
dc.identifier.issue1en_HK
dc.identifier.spage33en_HK
dc.identifier.epage39en_HK
dc.identifier.eissn1090-2104-
dc.identifier.isiWOS:000243336900006-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridHuen, MSY=23004751500en_HK
dc.identifier.scopusauthoridLu, LY=8686996700en_HK
dc.identifier.scopusauthoridLiu, DP=21934191400en_HK
dc.identifier.scopusauthoridHuang, JD=8108660600en_HK
dc.identifier.issnl0006-291X-

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