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

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

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

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
JournalNature Biotechnology
Publication statusPublished - 29 Jun 2015

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