CRISPR-Based Self-Cleaving Mechanism for Controllable Gene Delivery in Human Cells

dc.contributor.ISNI0000 0001 2535 9739 (Bleris, L)en_US
dc.contributor.LCNA2012076942‏ (Bleris, L)en_US
dc.contributor.authorMoore, Richarden_US
dc.contributor.authorSpinhirne, Alecen_US
dc.contributor.authorLai, Michael J.en_US
dc.contributor.authorPreisser, Samanthaen_US
dc.contributor.authorLi, Yien_US
dc.contributor.authorKang, Taeken_US
dc.contributor.authorBleris, Leonidasen_US
dc.descriptionSupplementary materials are available at Nucleic Acids Research Online (see DOI).en_US
dc.description.abstractControllable gene delivery via vector-based systems remains a formidable challenge in mammalian synthetic biology and a desirable asset in gene therapy applications. Here, we introduce a methodology to control the copies and residence time of a gene product delivered in host human cells but also selectively disrupt fragments of the delivery vehicle. A crucial element of the proposed system is the CRISPR protein Cas9. Upon delivery, Cas9 guided by a custom RNA sequence cleaves the delivery vector at strategically placed targets thereby inactivating a co-expressed gene of interest. Importantly, using experiments in human embryonic kidney cells, we show that specific parameters of the system can be adjusted to fine-tune the delivery properties. We envision future applications in complex synthetic biology architectures, gene therapy and trace-free delivery.;en_US
dc.description.sponsorshipUS National Institutes of Health (GM098984, GM096271, CA17001801); US National Science Foundation (CBNET-1105524)en_US
dc.identifier.bibliographicCitationMoore, Richard, Alec Spinhime, Michael J. Lai, Samantha Preisser, et al. 2014. "CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells." Nucleic Acids Research 43(2): doi:10.1093/nar/gku1326.
dc.publisherOxford University Pressen_US
dc.rightsCC BY-NC 4.0 (Attribution-Non-commercial)en_US
dc.rights©2014 The Authorsen_US
dc.source.journalNucleic Acids Researchen_US
dc.subjectmKate2 proteinen_US
dc.subjectCas9 proteinen_US
dc.subjectClustered Regularly Interspaced Short Palindromic Repeats (CRISPR)en_US
dc.subjectGuide RNA (gRNA)en_US
dc.subjectGene deliveryen_US
dc.titleCRISPR-Based Self-Cleaving Mechanism for Controllable Gene Delivery in Human Cellsen_US


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