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

Ayal Hendel, Rasmus O. Bak, Joseph T. Clark, Andrew B. Kennedy, Daniel E. Ryan, Subhadeep Roy, Israel Steinfeld, Benjamin D. Lunstad, Robert J. Kaiser, Alec B. Wilkens, Rosa Bacchetta, Anya Tsalenko, Douglas Dellinger, Laurakay Bruhn, Matthew H. Porteus

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)985-989
Number of pages5
JournalNature Biotechnology
Volume33
Issue number9
DOIs
Publication statusPublished - Sep 10 2015

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Biotechnology
  • Molecular Medicine
  • Bioengineering
  • Biomedical Engineering

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