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

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–...

Full description

Saved in:
Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2016-10, Vol.17 (19), p.1809-1812
Main Authors: He, Kaizhang, Chou, Eldon T., Begay, Shawn, Anderson, Emily M., van Brabant Smith, Anja
Format: Article
Language:English
Subjects:
Communication
Communications
conjugation
CRISPR-Cas Systems - genetics
CRISPR-Cas9
Editing
Gene Editing
gene technology
Molecular Structure
oligonucleotides
RNA, Guide, CRISPR-Cas Systems - chemistry
RNA, Guide, CRISPR-Cas Systems - metabolism
single guide RNA
Triazoles - chemistry
Triazoles - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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. Editing efficiency: A triazole‐linked single guide RNA (sgRNA) was constructed by using click chemistry. The sgRNA showed efficient CRISPR‐Cas9 genome editing activity comparable to that of natural dual RNA and unmodified sgRNA constructs. These findings open new avenues for generating long sgRNA libraries in array or plate formats.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201600320