Format

Send to

Choose Destination
Science. 2017 Nov 24;358(6366):1019-1027. doi: 10.1126/science.aaq0180. Epub 2017 Oct 25.

RNA editing with CRISPR-Cas13.

Cox DBT1,2,3,4,5,6, Gootenberg JS1,2,3,4,7, Abudayyeh OO1,2,3,4,6, Franklin B1,2,3,4, Kellner MJ1,2,3,4, Joung J1,2,3,4, Zhang F8,2,3,4.

Author information

1
Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.
2
McGovern Institute for Brain Research, MIT, Cambridge, MA 02139, USA.
3
Department of Brain and Cognitive Science, MIT, Cambridge, MA 02139, USA.
4
Department of Biological Engineering, MIT, Cambridge, MA 02139, USA.
5
Department of Biology, MIT, Cambridge, MA 02139, USA.
6
Harvard-MIT Division of Health Sciences and Technology, MIT, Cambridge, MA 02139, USA.
7
Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
8
Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA. zhang@broadinstitute.org.

Abstract

Nucleic acid editing holds promise for treating genetic disease, particularly at the RNA level, where disease-relevant sequences can be rescued to yield functional protein products. Type VI CRISPR-Cas systems contain the programmable single-effector RNA-guided ribonuclease Cas13. We profiled type VI systems in order to engineer a Cas13 ortholog capable of robust knockdown and demonstrated RNA editing by using catalytically inactive Cas13 (dCas13) to direct adenosine-to-inosine deaminase activity by ADAR2 (adenosine deaminase acting on RNA type 2) to transcripts in mammalian cells. This system, referred to as RNA Editing for Programmable A to I Replacement (REPAIR), which has no strict sequence constraints, can be used to edit full-length transcripts containing pathogenic mutations. We further engineered this system to create a high-specificity variant and minimized the system to facilitate viral delivery. REPAIR presents a promising RNA-editing platform with broad applicability for research, therapeutics, and biotechnology.

PMID:
29070703
PMCID:
PMC5793859
DOI:
10.1126/science.aaq0180
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center