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dc.contributor.authorSander, Jeffry D
dc.contributor.authorZaback, Peter
dc.contributor.authorJoung, Jae Keith
dc.contributor.authorVoytas, Daniel F.
dc.contributor.authorDobbs, Drena
dc.date.accessioned2011-04-28T04:22:53Z
dc.date.issued2009
dc.identifier.citationSander, Jeffry D., Peter Zaback, J. Keith Joung, Daniel F. Voytas, and Drena Dobbs. 2009. An affinity-based scoring scheme for predicting DNA-binding activities of modularly assembled zinc-finger proteins. Nucleic Acids Research 37(2): 506-515.en_US
dc.identifier.issn0305-1048en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:4878923
dc.description.abstractZinc-finger proteins (ZFPs) have long been recognized for their potential to manipulate genetic information because they can be engineered to bind novel DNA targets. Individual zinc-finger domains (ZFDs) bind specific DNA triplet sequences; their apparent modularity has led some groups to propose methods that allow virtually any desired DNA motif to be targeted in vitro. In practice, however, ZFPs engineered using this ‘modular assembly’ approach do not always function well in vivo. Here we report a modular assembly scoring strategy that both identifies combinations of modules least likely to function efficiently in vivo and provides accurate estimates of their relative binding affinities in vitro. Predicted binding affinities for 53 ‘three-finger’ ZFPs, computed based on energy contributions of the constituent modules, were highly correlated (r = 0.80) with activity levels measured in bacterial two-hybrid assays. Moreover, Kd values for seven modularly assembled ZFPs and their intended targets, measured using fluorescence anisotropy, were also highly correlated with predictions (r = 0.91). We propose that success rates for ZFP modular assembly can be significantly improved by exploiting the score-based strategy described here.en_US
dc.language.isoen_USen_US
dc.publisherOxford University Pressen_US
dc.relation.isversionofdoi:10.1093/nar/gkn962en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632909/pdf/en_US
dash.licenseLAA
dc.titleAn affinity-based scoring scheme for predicting DNA-binding activities of modularly assembled zinc-finger proteinsen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalNucleic Acids Researchen_US
dash.depositing.authorJoung, Jae Keith
dc.date.available2011-04-28T04:22:53Z
dash.affiliation.otherHMS^Pathologyen_US
dc.identifier.doi10.1093/nar/gkn962*
dash.contributor.affiliatedJoung, Keith
dash.contributor.affiliatedSander, Jeffry D.


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