Boosting plant genome editing with a versatile CRISPR-Combo system
Boosting plant genome editing with a versatile CRISPR-Combo system
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DOI：
10.1038/s41477-022-01151-9
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发表时间：
2022-05-01
期刊：
影响因子：
18
通讯作者：
Qi, Yiping
中科院分区：
文献类型：
--
作者：
Pan, Changtian;Li, Gen;Qi, Yiping
潘等人开发了一个通用的CRISPR-Combo平台，用于同时进行植物基因组编辑(靶向突变或碱基编辑)和基因激活，这是一种通用的基因组工程工具，在作物育种中具有广阔的应用前景。CRISPR-Cas9及其衍生的碱基编辑程序和CRISPR激活系统极大地辅助了植物基因组工程。然而，这些系统大多是单独使用的，它们的组合潜力在很大程度上尚未开发。在这里，我们开发了一个通用的CRISPR-Combo平台，基于单个Cas9蛋白，用于同时进行基因组编辑(靶向突变或碱基编辑)和植物基因激活。我们展示了CRISPR-Combo在促进植物基因组编辑方面的强大应用程序。首先，CRISPR-Combo被用来通过激活拟南芥中的成花素基因来缩短植物的生命周期，减少筛选无转基因基因组编辑植物的工作量。接下来，我们展示了通过激活杨树中的形态发生基因来加速基因组编辑植物的再生和繁殖。此外，我们还利用CRISPR-Combo技术实现了水稻不含外源植物激素的再生，这是一种主要丰富可遗传靶向突变的新方法。总之，CRISPR-COMBO是一种通用的基因组工程工具，在作物育种中具有广阔的应用前景。
Pan et al. develop a versatile CRISPR-Combo platform for simultaneous genome editing (targeted mutagenesis or base editing) and gene activation in plants, representing a versatile genome engineering tool with promising applications in crop breeding.CRISPR-Cas9, its derived base editors and CRISPR activation systems have greatly aided genome engineering in plants. However, these systems are mostly used separately, leaving their combinational potential largely untapped. Here we develop a versatile CRISPR-Combo platform, based on a single Cas9 protein, for simultaneous genome editing (targeted mutagenesis or base editing) and gene activation in plants. We showcase the powerful applications of CRISPR-Combo for boosting plant genome editing. First, CRISPR-Combo is used to shorten the plant life cycle and reduce the efforts in screening transgene-free genome-edited plants by activation of a florigen gene in Arabidopsis. Next, we demonstrate accelerated regeneration and propagation of genome-edited plants by activation of morphogenic genes in poplar. Furthermore, we apply CRISPR-Combo to achieve rice regeneration without exogenous plant hormones, which is established as a new method to predominately enrich heritable targeted mutations. In conclusion, CRISPR-Combo is a versatile genome engineering tool with promising applications in crop breeding.