title: Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing.
abstract: The CRISPR-Cas9 gene editing system requires Cas9 endonuclease and guide RNAs (either the natural dual RNA consisting of crRNA and tracrRNA or a chimeric single guide RNA) that direct site-specific double-stranded DNA cleavage. This communication describes a click ligation approach that uses alkyne-azide cycloaddition to generate a triazole-linked single guide RNA (sgRNA). The conjugated sgRNA shows efficient and comparable genome editing activity to natural dual RNA and unmodified sgRNA constructs.
authors.affiliation: GE Healthcare Dharmacon Inc., 2650 Crescent Drive, Suite 100, Lafayette, CO, 80026, USA.
authors.forename: Kaizhang
authors.initials: K
authors.lastname: He
authors.affiliation: GE Healthcare Dharmacon Inc., 2650 Crescent Drive, Suite 100, Lafayette, CO, 80026, USA.
authors.forename: Eldon T
authors.initials: ET
authors.lastname: Chou
authors.affiliation: GE Healthcare Dharmacon Inc., 2650 Crescent Drive, Suite 100, Lafayette, CO, 80026, USA.
authors.forename: Shawn
authors.initials: S
authors.lastname: Begay
authors.affiliation: GE Healthcare Dharmacon Inc., 2650 Crescent Drive, Suite 100, Lafayette, CO, 80026, USA.
authors.forename: Emily M
authors.initials: EM
authors.lastname: Anderson
authors.affiliation: GE Healthcare Dharmacon Inc., 2650 Crescent Drive, Suite 100, Lafayette, CO, 80026, USA. anja.smith@ge.com.
authors.forename: Anja
authors.initials: A
authors.lastname: van Brabant Smith
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D000072669:Gene Editing; D015394:Molecular Structure; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000737:chemistry / Q000378:metabolism*; D014230:Triazoles / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D014230:Triazoles
keywords: CRISPR-Cas9; conjugation; gene technology; oligonucleotides; single guide RNA