Issue 26, 2022

Cas9-mediated DNA cleavage guided by enzymatically prepared 4′-thio-modified RNA

Abstract

CRISPR-Cas9-mediated DNA editing relies on guide RNAs (gRNAs) that direct site-specific DNA cleavage by the Cas endonuclease. Because natural gRNA is susceptible to intracellular degradation, it is desirable to chemically protect it for efficient editing. Using 4′-thioribonucleoside 5′-triphosphates and T7 transcription, we have prepared 4′-thio-modified gRNAs that guide Cas9-mediated DNA cleavage. This approach is a simple way to obtain chemically modified RNA suitable for CRISPR-Cas9 DNA editing.

Graphical abstract: Cas9-mediated DNA cleavage guided by enzymatically prepared 4′-thio-modified RNA

Supplementary files

Article information

Article type
Communication
Submitted
20 Apr. 2022
Accepted
14 Jūn. 2022
First published
16 Jūn. 2022

Org. Biomol. Chem., 2022,20, 5245-5248

Cas9-mediated DNA cleavage guided by enzymatically prepared 4′-thio-modified RNA

N. Saito-Tarashima, M. Ueno, A. Murai, A. Matsuo and N. Minakawa, Org. Biomol. Chem., 2022, 20, 5245 DOI: 10.1039/D2OB00742H

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