Spelling Changes and Fluorescent Tagging With Prime Editing Vectors for Plants.
Spelling Changes and Fluorescent Tagging With Prime Editing Vectors for Plants.
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DOI：
10.3389/fgeed.2021.617553
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发表时间：
2021
影响因子：
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通讯作者：
Bogdanove AJ
中科院分区：
文献类型：
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作者：
Wang L;Kaya HB;Zhang N;Rai R;Willmann MR;Carpenter SCD;Read AC;Martin F;Fei Z;Leach JE;Martin GB;Bogdanove AJ
引物编辑是CRISPR-Cas系统的适应，其使用Cas9（H840 A）-逆转录酶融合物和用模板和引物结合位点序列修改的指导RNA以实现靶DNA的RNA模板化转化，从而允许指定的取代、插入和缺失。在第一份关于植物中prime编辑的报告中，描述了水稻和小麦中的各种编辑，包括高达15 bp的插入。几项水稻研究很快跟进，但没有一个报道更大的插入。在这里，我们报告了易于使用的载体，用于双子叶植物和单子叶植物中的主要编辑，它们在本氏烟草、水稻和拟南芥中的验证，以及66 bp的插入，该插入使分裂GFP荧光标记成为可能。
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 Nicotiana benthamiana, rice, and Arabidopsis, and an insertion of 66 bp that enabled split-GFP fluorescent tagging.