Conjugation and Evaluation of Triazole-Linked Single Guide RNA for CRISPR-Cas9 Gene Editing

被引:25
作者
He, Kaizhang [1 ]
Chou, Eldon T. [1 ]
Begay, Shawn [1 ]
Anderson, Emily M. [1 ]
Smith, Anja van Brabant [1 ]
机构
[1] GE Healthcare Dharmacon Inc, 2650 Crescent Dr,Suite 100, Lafayette, CO 80026 USA
关键词
conjugation; CRISPR-Cas9; gene technology; oligonucleotides; single guide RNA; SOLID-PHASE; HUMAN-CELLS; GENOME; CAS9; CHEMISTRY; ENDONUCLEASE; DELIVERY; SYSTEMS; DNA;
D O I
10.1002/cbic.201600320
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:1809 / 1812
页数:4
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