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CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells.

ABSTRACT: 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.

SUBMITTER: Moore R

PROVIDER: S-EPMC4333380 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells.

Moore Richard R Spinhirne Alec A Lai Michael J MJ Preisser Samantha S Li Yi Y Kang Taek T Bleris Leonidas L

Nucleic acids research 20141218 2

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 ...[more]

PMID: 25527740

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