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dc.contributor.authorYang, Luhanen_US
dc.contributor.authorGuell, Marcen_US
dc.contributor.authorByrne, Susanen_US
dc.contributor.authorYang, Joyce L.en_US
dc.contributor.authorDe Los Angeles, Alejandroen_US
dc.contributor.authorMali, Prashanten_US
dc.contributor.authorAach, Johnen_US
dc.contributor.authorKim-Kiselak, Carolineen_US
dc.contributor.authorBriggs, Adrian Wen_US
dc.contributor.authorRios, Xavieren_US
dc.contributor.authorHuang, Po-Yien_US
dc.contributor.authorDaley, Georgeen_US
dc.contributor.authorChurch, Georgeen_US
dc.date.accessioned2014-03-10T20:32:43Z
dc.date.issued2013en_US
dc.identifier.citationYang, L., M. Guell, S. Byrne, J. L. Yang, A. De Los Angeles, P. Mali, J. Aach, et al. 2013. “Optimization of scarless human stem cell genome editing.” Nucleic Acids Research 41 (19): 9049-9061. doi:10.1093/nar/gkt555. http://dx.doi.org/10.1093/nar/gkt555.en
dc.identifier.issn0305-1048en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:11878811
dc.description.abstractEfficient strategies for precise genome editing in human-induced pluripotent cells (hiPSCs) will enable sophisticated genome engineering for research and clinical purposes. The development of programmable sequence-specific nucleases such as Transcription Activator-Like Effectors Nucleases (TALENs) and Cas9-gRNA allows genetic modifications to be made more efficiently at targeted sites of interest. However, many opportunities remain to optimize these tools and to enlarge their spheres of application. We present several improvements: First, we developed functional re-coded TALEs (reTALEs), which not only enable simple one-pot TALE synthesis but also allow TALE-based applications to be performed using lentiviral vectors. We then compared genome-editing efficiencies in hiPSCs mediated by 15 pairs of reTALENs and Cas9-gRNA targeting CCR5 and optimized ssODN design in conjunction with both methods for introducing specific mutations. We found Cas9-gRNA achieved 7–8× higher non-homologous end joining efficiencies (3%) than reTALENs (0.4%) and moderately superior homology-directed repair efficiencies (1.0 versus 0.6%) when combined with ssODN donors in hiPSCs. Using the optimal design, we demonstrated a streamlined process to generated seamlessly genome corrected hiPSCs within 3 weeks.en
dc.language.isoen_USen
dc.publisherOxford University Pressen
dc.relation.isversionofdoi:10.1093/nar/gkt555en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3799423/pdf/en
dash.licenseLAAen_US
dc.titleOptimization of scarless human stem cell genome editingen
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalNucleic Acids Researchen
dash.depositing.authorYang, Luhanen_US
dc.date.available2014-03-10T20:32:43Z
dc.identifier.doi10.1093/nar/gkt555*
dash.authorsorderedfalse
dash.contributor.affiliatedDe Los Angeles, Alejandro
dash.contributor.affiliatedYang, Joyce
dash.contributor.affiliatedHuang, Po-Yi
dash.contributor.affiliatedGuell, Marc
dash.contributor.affiliatedBriggs, Adrian
dash.contributor.affiliatedYang, Luhan
dash.contributor.affiliatedMali, Prashant
dash.contributor.affiliatedByrne, Susan M
dash.contributor.affiliatedDaley, George
dash.contributor.affiliatedAach, John
dash.contributor.affiliatedChurch, George


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