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

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

DOI

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

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.

OriginalsprogEngelsk
TidsskriftNature Biotechnology
ISSN1087-0156
DOI
StatusUdgivet - 29 jun. 2015

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