Journal
CHEMBIOCHEM
Volume 17, Issue 19, Pages 1809-1812Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201600320
Keywords
conjugation; CRISPR-Cas9; gene technology; oligonucleotides; single guide RNA
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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.
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