Format

Send to

Choose Destination
Nucleic Acids Res. 2015 Jan;43(2):1297-303. doi: 10.1093/nar/gku1326. Epub 2014 Dec 18.

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

Author information

1
Bioengineering Department, University of Texas at Dallas, Richardson, TX 75080, USA Electrical Engineering Department, University of Texas at Dallas, Richardson, TX 75080, USA.
2
Bioengineering Department, University of Texas at Dallas, Richardson, TX 75080, USA.
3
Center for Systems Biology, University of Texas at Dallas, Richardson, TX 75080, USA.
4
Department of Molecular & Cell Biology, University of Texas at Dallas, Richardson, TX 75080, USA.
5
Bioengineering Department, University of Texas at Dallas, Richardson, TX 75080, USA Department of Chemistry, University of Texas at Dallas, Richardson, TX 75080, USA Electrical Engineering Department, University of Texas at Dallas, Richardson, TX 75080, USA bleris@utdallas.edu.

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.

PMID:
25527740
PMCID:
PMC4333380
DOI:
10.1093/nar/gku1326
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Silverchair Information Systems Icon for PubMed Central
Loading ...
Support Center