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.

CRISPR-Cas9、其衍生的碱基编辑器和 CRISPR 激活系统极大地帮助了植物的基因组工程。然而，这些系统大多单独使用，使得它们的组合潜力在很大程度上未被开发。在这里，我们开发了一个基于单一 Cas9 蛋白的多功能 CRISPR-Combo 平台，用于在植物中同时进行基因组编辑（靶向诱变或碱基编辑）和基因激活。我们展示了 CRISPR-Combo 在促进植物基因组编辑方面的强大应用。首先，CRISPR-Combo 用于通过激活拟南芥中的成花基因来缩短植物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，我们应用 CRISPR-Combo 在没有外源植物激素的情况下实现水稻再生，这是一种主要丰富可遗传靶向突变的新方法。总之，CRISPR-Combo 是一种多功能的基因组工程工具，在作物育种中具有广阔的应用前景。