title: Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells.
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.
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Ayal
authors.initials: A
authors.lastname: Hendel
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Rasmus O
authors.initials: RO
authors.lastname: Bak
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Joseph T
authors.initials: JT
authors.lastname: Clark
authors.affiliation: Agilent Research Laboratories, Santa Clara, California, USA.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Kennedy
authors.affiliation: Agilent Research Laboratories, Santa Clara, California, USA.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Ryan
authors.affiliation: Agilent Research Laboratories, Boulder, Colorado, USA.
authors.forename: Subhadeep
authors.initials: S
authors.lastname: Roy
authors.affiliation: Agilent Research Laboratories, Tel Aviv, Israel.
authors.forename: Israel
authors.initials: I
authors.lastname: Steinfeld
authors.affiliation: Agilent Research Laboratories, Boulder, Colorado, USA.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Lunstad
authors.affiliation: Agilent Research Laboratories, Santa Clara, California, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Kaiser
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Alec B
authors.initials: AB
authors.lastname: Wilkens
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Rosa
authors.initials: R
authors.lastname: Bacchetta
authors.affiliation: Agilent Research Laboratories, Santa Clara, California, USA.
authors.forename: Anya
authors.initials: A
authors.lastname: Tsalenko
authors.affiliation: Agilent Research Laboratories, Boulder, Colorado, USA.
authors.forename: Douglas
authors.initials: D
authors.lastname: Dellinger
authors.affiliation: Agilent Research Laboratories, Santa Clara, California, USA.
authors.forename: Laurakay
authors.initials: L
authors.lastname: Bruhn
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Porteus
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000737:chemistry* / Q000235:genetics*
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems