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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Moore R,Spinhirne A,Lai MJ,Preisser S,Li Y,Kang T,Bleris L

doi

10.1093/nar/gku1326

subject

Has Abstract

pub_date

2015-01-01 00:00:00

pages

1297-303

issue

2

eissn

0305-1048

issn

1362-4962

pii

gku1326

journal_volume

43

pub_type

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