Bellagen Biotechnology Co. Ltd, Ji'nan, China.
School of Life Sciences, Shandong Normal University, Ji'nan, China.
Plant Biotechnol J. 2024 Feb;22(2):379-385. doi: 10.1111/pbi.14192. Epub 2023 Oct 11.
The CRISPR/Cas type V-I is a family of programmable nuclease systems that prefers a T-rich protospacer adjacent motif (PAM) and is guided by a short crRNA. In this study, the genome-editing application of Cas12i3, a type V-I family endonuclease, was characterized in rice. We developed a CRIPSR/Cas12i3-based Multiplex direct repeats (DR)-spacer Array Genome Editing (iMAGE) system that was efficient in editing various genes in rice. Interestingly, iMAGE produced chromosomal structural variations with a higher frequency than CRISPR/Cas9. In addition, we developed base editors using deactivated Cas12i3 and generated herbicide-resistant rice plants using the base editors. These CRIPSR/Cas12i3-based genome editing systems will facilitate precision molecular breeding in plants.
CRISPR/Cas 类型 V-I 是一类可编程核酸内切酶系统，其偏好富含 T 的原间隔序列邻近基序 (PAM)，并由短的 crRNA 引导。在这项研究中，我们对 Cas12i3（一种 V-I 家族内切酶）在水稻中的基因组编辑应用进行了研究。我们开发了一种基于 CRISPR/Cas12i3 的多聚重复直接重复（DR）-间隔区阵列基因组编辑（iMAGE）系统，该系统可有效地编辑水稻中的各种基因。有趣的是，iMAGE 产生的染色体结构变异的频率高于 CRISPR/Cas9。此外，我们使用失活的 Cas12i3 开发了碱基编辑器，并使用碱基编辑器生成了抗除草剂的水稻植株。这些基于 CRISPR/Cas12i3 的基因组编辑系统将有助于在植物中进行精确的分子育种。
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