Moore, RichardSpinhirne, AlecLai, Michael J.Preisser, SamanthaLi, YiKang, TaekBleris, Leonidas2015-03-302015-03-302014-12-182014-12-181362-4962http://hdl.handle.net/10735.1/4396Supplementary materials are available at Nucleic Acids Research Online (see DOI).Controllable 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.;CC BY-NC 4.0 (Attribution-Non-commercial)©2014 The Authorshttp://creativecommons.org/licenses/by-nc/4.0/mKate2 proteinCas9 proteinClustered Regularly Interspaced Short Palindromic Repeats (CRISPR)Guide RNA (gRNA)Gene deliveryarticleMoore, 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.