Show simple item record

dc.contributor.authorHowden, SE
dc.contributor.authorMaufort, JP
dc.contributor.authorDuffin, BM
dc.contributor.authorElefanty, AG
dc.contributor.authorStanley, EG
dc.contributor.authorThomson, JA
dc.date.accessioned2021-02-05T01:04:57Z
dc.date.available2021-02-05T01:04:57Z
dc.date.issued2015-12-08
dc.identifierpii: S2213-6711(15)00306-9
dc.identifier.citationHowden, S. E., Maufort, J. P., Duffin, B. M., Elefanty, A. G., Stanley, E. G. & Thomson, J. A. (2015). Simultaneous Reprogramming and Gene Correction of Patient Fibroblasts.. Stem Cell Reports, 5 (6), pp.1109-1118. https://doi.org/10.1016/j.stemcr.2015.10.009.
dc.identifier.issn2213-6711
dc.identifier.urihttp://hdl.handle.net/11343/260263
dc.description.abstractThe derivation of genetically modified induced pluripotent stem (iPS) cells typically involves multiple steps, requiring lengthy cell culture periods, drug selection, and several clonal events. We report the generation of gene-targeted iPS cell lines following a single electroporation of patient-specific fibroblasts using episomal-based reprogramming vectors and the Cas9/CRISPR system. Simultaneous reprogramming and gene targeting was tested and achieved in two independent fibroblast lines with targeting efficiencies of up to 8% of the total iPS cell population. We have successfully targeted the DNMT3B and OCT4 genes with a fluorescent reporter and corrected the disease-causing mutation in both patient fibroblast lines: one derived from an adult with retinitis pigmentosa, the other from an infant with severe combined immunodeficiency. This procedure allows the generation of gene-targeted iPS cell lines with only a single clonal event in as little as 2 weeks and without the need for drug selection, thereby facilitating "seamless" single base-pair changes.
dc.languageeng
dc.publisherElsevier BV
dc.titleSimultaneous Reprogramming and Gene Correction of Patient Fibroblasts.
dc.typeJournal Article
dc.identifier.doi10.1016/j.stemcr.2015.10.009
melbourne.affiliation.departmentPaediatrics (RCH)
melbourne.affiliation.facultyMedicine, Dentistry & Health Sciences
melbourne.source.titleStem Cell Reports
melbourne.source.volume5
melbourne.source.issue6
melbourne.source.pages1109-1118
dc.rights.licenseCC BY-NC-ND
melbourne.elementsid1046382
melbourne.openaccess.pmchttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682122
melbourne.contributor.authorElefanty, Andrew
melbourne.contributor.authorStanley, Edouard
dc.identifier.eissn2213-6711
melbourne.accessrightsOpen Access


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record