MS2 RNA aptamer enhances prime editing in rice

作者：Yiping Chai, Yuanyuan Jiang, Junya Wang, Dexin Qiao, Yu Zhang, Cuiping Xin, Yun Zhou, Xuechen Wang, Qijun Chen · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2021 · DOI：10.1101/2021.10.20.465209 · 被引用次数：7 · 研究领域：CRISPR and Genetic Engineering、RNA regulation and disease、RNA and protein synthesis mechanisms

Abstract Prime editing is a universal and very promising precise genome editing technology. However, optimization of prime editor (PE) from different aspects remains vital for its use as a routine tool in plant basic research and crop molecular breeding. In this report, we tested MS2-based prime editor (MS2PE). We fused the M-MLV reverse transcriptase (RT) gene variant to the MS2 RNA binding protein gene, MCP , and allowed the MCP-RT fusion gene to co-express with the SpCas9 nickase gene, SpCas9H840A , and various engineered pegRNAs harboring MS2 RNA (MS2pegR). Compared with control PEs, MS2PEs significantly enhanced editing efficiency at four of six targets in rice protoplasts, and achieved 1.2∼10.1-fold increase in editing efficiency at five of six targets in transgenic rice lines. Furthermore, we tested total 22 different MS2pegR scaffolds, 3 RT variants or genes, 2 MCP variants, and various combinations of the Cas9 nickase, RT, and MCP modules. Our results demonstrated an alternative strategy for enhancing prime editing.