Mengyuan Liu, Xiang Zhang, Wen Xu, Guiting Kang, Ya Liu, Xinxiang Liu, Wen Ren, Jiuran Zhao, Jinxiao Yang
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引用次数: 0
摘要
高效、精确的基因组缺失为植物研究中的蛋白质功能调查和提高农业性状带来了希望。在这项研究中,我们测试了 PRIME-Del (PDel) 策略,该策略使用一对以相反 DNA 链为目标的质粒编辑向导 RNA(pegRNA),在 6 个内源靶点的 60 bp 片段缺失中实现了 55.8% 的平均缺失效率。此外,在转基因水稻植株中,对 OsGS1 位点 2000 bp 片段的精确删除效率高达 84.2%。为了添加两个核酸序列之间被无意删除的碱基,我们采用了 PDel/Syn 策略,在 RT 模板中需要修补的区域引入多个同义碱基突变。PDel/Syn策略在六个内源性靶点平均实现了58.1%的删除效率,高于PDel策略。本研究提出的策略有助于在转基因水稻植物中实现更准确、更灵活的删除:在线版本包含补充材料,可查阅 10.1007/s42994-024-00153-9。
Efficient and precise genomic deletion in rice using enhanced prime editing
Efficient and precise genomic deletion shows promise for investigating the function of proteins in plant research and enhancing agricultural traits. In this study, we tested the PRIME-Del (PDel) strategy using a pair of prime editing guide RNAs (pegRNAs) that targeted opposite DNA strands and achieved an average deletion efficiency of 55.8% for 60 bp fragment deletions at six endogenous targets. Moreover, as high as 84.2% precise deletion efficiency was obtained for a 2000 bp deletion at the OsGS1 site in transgenic rice plants. To add the bases that were unintentionally deleted between the two nicking sequences, we used the PDel/Syn strategy, which introduced multiple synonymous base mutations in the region that had to be patched in the RT template. The PDel/Syn strategy achieved an average of 58.1% deletion efficiency at six endogenous targets, which was higher than the PDel strategy. The strategies presented in this study contribute to achieving more accurate and flexible deletions in transgenic rice plants.