Development of PAM-flexible multiplex genome editors for enhancing cold-tolerance in indica rice

文献类型： 外文期刊

第一作者： Tan, Jiantao

作者： Tan, Jiantao;Sun, Kangli;Qian, Qian;Peng, Xin;Shen, Mengyuan;Zhou, Degui;Liu, Qi;Chai, Nan;Liu, Yao-Guang;Zhu, Qinlong

作者机构：

关键词： Genetic engineering; Genetic transformation; Genome editing; WRKY transcription factors

期刊名称：CROP JOURNAL （ 影响因子：5.6； 五年影响因子：6.0 ）

ISSN： 2095-5421

年卷期： 2025 年 13 卷 3 期

页码：

收录情况： SCI

摘要： The canonical CRISPR/Cas editors are constrained by the requirement for specific PAMs, which substantially limits their editable target range. Rice (particularly indica rice) is sensitive to low temperature, which impacts the yield and restricts the geographic distribution of rice. In this study, we developed PAM-flexible multiplex genome editing tools based on SpG (recognising NGN-PAMs) and SpRY (recognising NNN-PAMs) variants. We then tested the feasibility of using a sweet potato leaf curl virus (SPLCV) replicon-based expression vector and single-stranded DNA-binding domain (DBD) to improve the editing efficiency of these PAM-flexible editors. Furthermore, we used SpG-mediated multiplex genome editing to achieve comprehensive improvement in cold tolerance in indica rice by editing WRKY transcription factors OsWRKY53 and OsWRKY63, to generate high cold-resistant indica rice lines. We concluded that these PAM-flexible multiplex genome editors are powerful tools for multi-gene editing for crop genetic improvement. (c) 2025 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NCND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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