Developing an efficient and visible prime editing system to restore tobacco 8-hydroxy-copalyl diphosphate gene for labdane diterpene Z-abienol biosynthesis  

作　　者：Jianduo Zhang Lu Zhang Chengwei Zhang Yongxing Yang Huayin Liu Lu Li Shengxue Zhang Xianggan Li Xinxiang Liu Ya Liu Jin Wang Guangyu Yang Qingyou Xia Weiguang Wang Jinxiao Yang 

机构地区：[1]Yunnan Key Laboratory of Tobacco,Yunnan Academy of Tobacco Science,Kunming 650231,China [2]Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding,Beijing Academy of Agriculture&Forestry Sciences,Beijing 100089,China [3]Key Laboratory of Natural Products Synthetic Biology of Ethnic Medicinal Endophytes,State Ethnic Affairs Commission,Yunnan Minzu University,Kunming 650031,China [4]Cropedit Biotechnology Co.Ltd.,Beijing 102206,China [5]Biological Science Research Center,Southwest University,Chongqing 400716,China

出　　处：《Science China(Life Sciences)》2023年第12期2910-2921,共12页中国科学（生命科学英文版）

基　　金：supported by Beijing Scholars Program (BSP041);Financial Special Fund of Beijing Academy of Agriculture and Forestry Sciences (CZZJ202206);the key projects of YNZY (2022JY02);CNTC (110202101034,JY-11)。

摘　　要：Prime editing(PE)is a versatile CRISPR-Cas based precise genome-editing platform widely used to introduce a range of possible base conversions in various organisms.However,no PE systems have been shown to induce heritable mutations in tobacco,nor in any other dicot.In this study,we generated an efficient PE system in tobacco that not only introduced heritable mutations,but also enabled anthocyanin-based reporter selection of transgene-free T_(1) plants.This system was used to confer Zabienol biosynthesis in the allotetraploid tobacco cultivar HHDJY by restoring a G>T conversion in the NtCPS2 gene.High levels of Z-abienol were detected in the leaves of homozygous T_(1) plants at two weeks after topping.This study describes an advance in PE systems and expands genome-editing toolbox in tobacco,even in dicots,for use in basic research and molecular breeding.And restoring biosynthesis of Z-abienol in tobacco might provide an efficient way to obtain Z-abienol in plants.

关 键 词：prime editing Z-abienol biosynthesis visible marker CRISPR Cas9 TOBACCO 

分 类 号：Q78[生物学—分子生物学]