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利用多功能 CRISPR-Combo 系统增强植物基因组编辑。

Boosting plant genome editing with a versatile CRISPR-Combo system.

机构信息

Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA.

Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD, USA.

出版信息

Nat Plants. 2022 May;8(5):513-525. doi: 10.1038/s41477-022-01151-9. Epub 2022 May 20.

Abstract

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.

摘要

CRISPR-Cas9、其衍生的碱基编辑器和 CRISPR 激活系统极大地促进了植物的基因组工程。然而,这些系统大多是分开使用的,其组合潜力在很大程度上尚未开发。在这里,我们基于单个 Cas9 蛋白开发了一种通用的 CRISPR-Combo 平台,用于在植物中同时进行基因组编辑(靶向突变或碱基编辑)和基因激活。我们展示了 CRISPR-Combo 在增强植物基因组编辑方面的强大应用。首先,CRISPR-Combo 通过激活拟南芥中的开花素基因来缩短植物的生命周期并减少筛选无转基因基因组编辑植物的工作量。接下来,我们通过在杨树中激活形态发生基因来展示加速基因组编辑植物的再生和繁殖的应用。此外,我们将 CRISPR-Combo 应用于实现无需外源植物激素的水稻再生,这被确立为一种主要富集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,在作物育种中有广阔的应用前景。

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