Strategic Base Modifications Refine RNA Function and Reduce CRISPR-Cas9 Off-targets

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2025 Feb 18

PMID 39964477

Authors

Kaisong Zhang

Wei Shen

Yunting Zhao

Xinyan Xu

Xingyu Liu

Qianqian Qi

Siqi Huang

Tian Tian

Xiang Zhou

Affiliations

Soon will be listed here.

Abstract

In contrast to traditional RNA regulatory approaches that modify the 2'-OH group, this study explores strategic base modifications using 5-carboxylcytosine (ca5C). We developed a technique where ca5C is transformed into dihydrouracil via treatment with borane-pyridine complex or 2-picoline borane complex, leading to base mutations that directly impact RNA functionality. This innovative strategy effectively manages CRISPR-Cas9 system activities, significantly minimizing off-target effects. Our approach not only demonstrates a significant advancement in RNA manipulation but also offers a new method for the precise control of gene editing technologies, showcasing its potential for broad application in chemical biology.

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