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| 1 | Base editing in plants: Current status and challenges显示文摘Genome editing technologies have revolutionized the field of plant science by enabling targeted modification of plant genomes and are emerging as powerful tools for both plant gene functional analyses and crop improvement. Although homology-directed repair(HDR)is a feasible approach to achieve precise gene replacement and base substitution in some plant species, the dominance of the non-homologous end joining pathway and low efficiency of HDR in plant cells have limited its application. Base editing has emerged as an alternative tool to HDR-mediated replacement, facilitating precise editing of plant genome by converting one single base to another in a programmable manner without a doublestranded break and a donor repair template. In this review, we summarize the latest developments in base-editing technologies as well as their underlying mechanisms. We review current applications of these technologies in plant species. Finally, we address the challenges and future perspectives of this emerging technology in plants. | Sutar Suhas Bharat Shaoya Li Jingying Li Lei Yan Lanqin Xia | 2020 | The Crop Journal2020，8，3： | 8 |
| 2 | 基于Csy4与MCP的新型迷你基因组编辑系统的构建显示文摘MCP是MS2噬菌体的外壳蛋白,Csy4是一种参与CRISPR 1-F系统crRNA生成的小型蛋白,能够以较高的特异性识别并结合RNA。目前CRISPR/Cas等基因组编辑技术存在靶向核酸酶分子量大、脱靶率高、受PAM位点限制等问题,为解决上述问题,构建基于上述两种小型蛋白的新型迷你基因组编辑系统。本研究采用AlphaFold2预测MCP-FokI、FokI-MCP、Csy4-FokI和FokI-Csy4融合蛋白的结构,通过浸花法将MCP-FokI和FokI-MCP编辑载体分别转化拟南芥,利用拟南芥叶片注射的方法投送CLCrV介导的Csy4-FokI与FokI-Csy4编辑系统,提取拟南芥基因组DNA,通过HI-TOM高通量测序检测新型迷你基因组编辑系统的编辑能力。结果显示,融合蛋白中MCP、FokI和Csy4都各自保持着自身原有的三维结构,预示它们都能正常发挥彼此的功能。构建靶向敲除拟南芥CLA1基因的4个不同中间间隔区的双靶位点MCP-FokI和MCP-FokI植物表达载体,初步证明MCP-FokI和FokI-MCP均不能实现对靶基因的靶向编辑。构建靶向敲除拟南芥CLA1基因的7个CLCrV介导的不同中间间隔区的双靶位点Csy4-FokI编辑载体,其中CLCrV介导的Csy4-FokI编辑系统能够实现对靶基因的靶向编辑,但是突变类型均为碱基置换类型且编辑效率很低,而FokI-Csy4基因组编辑体系并未检测到编辑的发生。成功构建了Csy4-FokI新型迷你基因组编辑系统,为克服CRISPR/Cas基因组编辑技术存在的问题提供了一种新的解决方案。 | 邓嘉辉 雷建峰 赵燚 刘敏 胡子曜 尤扬子 邵武奎 柳建飞 刘晓东 | 2023 | 生物技术通报2023，39，10： | 0 |
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