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Nat Biotechnol. 2015 Sep;33(9):985-989. doi: 10.1038/nbt.3290. Epub 2015 Jun 29.

Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells.

Author information

1
Department of Pediatrics, Stanford University, Stanford, California, USA.
2
Agilent Research Laboratories, Santa Clara, California, USA.
3
Agilent Research Laboratories, Boulder, Colorado, USA.
4
Agilent Research Laboratories, Tel Aviv, Israel.
#
Contributed equally

Abstract

CRISPR-Cas-mediated genome editing relies on guide RNAs that direct site-specific DNA cleavage facilitated by the Cas endonuclease. Here we report that chemical alterations to synthesized single guide RNAs (sgRNAs) enhance genome editing efficiency in human primary T cells and CD34(+) hematopoietic stem and progenitor cells. Co-delivering chemically modified sgRNAs with Cas9 mRNA or protein is an efficient RNA- or ribonucleoprotein (RNP)-based delivery method for the CRISPR-Cas system, without the toxicity associated with DNA delivery. This approach is a simple and effective way to streamline the development of genome editing with the potential to accelerate a wide array of biotechnological and therapeutic applications of the CRISPR-Cas technology.

PMID:
26121415
PMCID:
PMC4729442
DOI:
10.1038/nbt.3290
[Indexed for MEDLINE]
Free PMC Article

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