Wang Li, Kaya Hilal Betul, Zhang Ning, Rai Rhitu, Willmann Matthew R, Carpenter Sara C D, Read Andrew C, Martin Federico, Fei Zhangjun, Leach Jan E, Martin Gregory B, Bogdanove Adam J
Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, United States.
Department of Bioengineering, Faculty of Engineering, Manisa Celal Bayar University, Manisa, Turkey.
Front Genome Ed. 2021 Mar 4;3:617553. doi: 10.3389/fgeed.2021.617553. eCollection 2021.
Prime editing is an adaptation of the CRISPR-Cas system that uses a Cas9(H840A)-reverse transcriptase fusion and a guide RNA amended with template and primer binding site sequences to achieve RNA-templated conversion of the target DNA, allowing specified substitutions, insertions, and deletions. In the first report of prime editing in plants, a variety of edits in rice and wheat were described, including insertions up to 15 bp. Several studies in rice quickly followed, but none reported a larger insertion. Here, we report easy-to-use vectors for prime editing in dicots as well as monocots, their validation in , rice, and Arabidopsis, and an insertion of 66 bp that enabled split-GFP fluorescent tagging.
引导编辑是对CRISPR-Cas系统的一种改造,它使用Cas9(H840A)-逆转录酶融合蛋白和一个经模板和引物结合位点序列修饰的引导RNA,以实现目标DNA的RNA模板化转化,从而实现特定的替换、插入和缺失。在植物引导编辑的首次报告中,描述了水稻和小麦中的多种编辑,包括长达15个碱基对的插入。随后很快有几项关于水稻的研究,但都没有报告更大的插入。在这里,我们报告了适用于双子叶植物和单子叶植物引导编辑的易用载体、它们在水稻和拟南芥中的验证,以及一个能够实现分裂GFP荧光标记的66个碱基对的插入。