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Abstract
背景回顾：CRISPR/Cas-derived base editor enables precise editing of target sites and has been widely used for basic research and crop genetic improvement.
提出问题：However, the editing efficiency of base editors at different targets varies greatly. Here, we develop a set of highly efficient base editors in cotton plants.
结果1-剪辑编辑器优化：GhABE8e, which is fused to conventional nCas9, exhibits 99.9% editing efficiency, compared to GhABE7.10 with 64.9%, and no off-target editing is detected. We further replace nCas9 with dCpf1, which recognizes TTTV PAM sequences, to broaden the range of the target site.
结果2.1-TFL1编辑种质创新：To explore the functional divergence of TERMINAL FLOWER 1 (TFL1), we edit the non-coding and coding regions of GhTFL1 with 26 targets to generate a comprehensive allelic population including 300 independent lines in cotton.
结果2.2-优异种质筛选：This allows hidden pleiotropic roles for GhTFL1 to be revealed and allows us to rapidly achieve directed domestication of cotton and create ideotype germplasm with moderate height, shortened fruiting branches, compact plant, and early-flowering. Further, by exploring the molecular mechanism of the GhTFL1L86P and GhTFL1K53G+S78G mutations, we find that the GhTFL1L86P mutation weakens the binding strength of the GhTFL1 to other proteins but does not lead to a complete loss of GhTFL1 function.
结论：This strategy provides an important technical platform and genetic information for the study and creation of ideal plant architecture.
摘 要
背景：CRISPR/Cas驱动的碱基编辑器能够精确编辑目标位点，被广泛用于基础研究和作物遗传改良。但是，碱基编辑器在不同位点的编辑效率差异很大。本文中，作者开发出了一系列适合棉花的高效碱基编辑器。
剪辑编辑器优化:作者通过将GhABE8e融合传统的nCas9获得的编辑效率为99.9%，而GhABE7.10仅为64.9%，并且没有检测到脱靶编辑事件。作者进一步用dCpf1替换了nCas9，dCpf1能够识别TTTV PAM序列，拓宽了目标位点的范围。
TFL1编辑新种质创制及优异种质选育：为了解析TFL1的功能分化，作者编辑了GhTFL1基因上非编码区和编码区的序列，基于26个靶向位点，作者共获得了一个包含300个独立株系的等位变异群体。这使得作者能够揭示GhTFL1基因潜藏的多效性作用，从而实现棉花的快速定向驯化，创制中等株高、果枝缩短、紧凑株形以及早花的理想棉花种质。此外，通过挖掘GhTFL1L86P和GhTFL1K53G+S78G突变体的分子机制，作者发现GhTFL1L86P突变减弱了其结合到其他蛋白上的能力，但是并不会完全丢失GhTFL1的功能。
结论：本策略为研究和创制植物理想株形提供了重要的技术平台和遗传信息。华中农业大学金双侠教授、张献龙院士和石河子大学聂新辉教授为共同通讯作者，华中农业大学博士后王冠英为第一作者。该研究获得国家杰出青年科学基金和中国博士后科学基金等项目支持。
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