Format

Send to

Choose Destination
Methods. 2017 May 15;121-122:9-15. doi: 10.1016/j.ymeth.2017.04.003. Epub 2017 Apr 12.

Genome editing via delivery of Cas9 ribonucleoprotein.

Author information

1
Innovative Genomics Institute, University of California, Berkeley, CA, United States.
2
Innovative Genomics Institute, University of California, Berkeley, CA, United States; Department of Molecular and Cell Biology, University of California, Berkeley, CA, United States.
3
Innovative Genomics Institute, University of California, Berkeley, CA, United States; Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT, United States.

Abstract

The CRISPR-Cas genome editing system is very powerful. The format of the CRISPR reagents and the means of delivery are often important factors in targeting efficiency. Delivery of recombinant Cas9 protein and guide RNA (gRNA) as a preformed ribonucleoprotein (RNP) complex has recently emerged as a powerful and general approach to genome editing. Here we outline methods to produce and deliver Cas9 RNPs. A donor DNA carrying desired sequence changes can also be included to program precise sequence introduction or replacement. RNP delivery limits exposure to genome editing reagents, reduces off-target events, drives high rates of homology-dependent repair, and can be applied to embryos to rapidly generate animal models. RNP delivery thus minimizes some of the pitfalls of alternative editing modalities and is rapidly being adopted by the genome editing community.

PMID:
28410976
DOI:
10.1016/j.ymeth.2017.04.003
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center