Department of Biotechnology, Center for Informational Biology, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, China.
Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA.
Methods Mol Biol. 2021;2238:193-204. doi: 10.1007/978-1-0716-1068-8_12.
CRISPR-Cas9 and Cas12a (formerly Cpf1), RNA-guided DNA endonucleases found from adaptive immune system in prokaryotes, have been engineered and widely adopted as two of the most powerful genome editing systems in plants. Recently, we developed a single transcript unit (STU) CRISPR 2.0 toolbox for applications in plants, which contains two STU-Cas9 systems and one STU-Cas12a system. Here, we describe a detailed protocol about using the STU CRISPR 2.0 systems to achieve single and multiplex genome editing in rice.
CRISPR-Cas9 和 Cas12a(以前称为 Cpf1)是原核生物适应性免疫系统中发现的 RNA 指导的 DNA 内切酶,已被工程化并广泛应用于植物的两种最强大的基因组编辑系统。最近,我们开发了一个单转录单元 (STU) CRISPR 2.0 工具包,用于植物应用,其中包含两个 STU-Cas9 系统和一个 STU-Cas12a 系统。在这里,我们描述了一个详细的方案,介绍如何使用 STU CRISPR 2.0 系统在水稻中实现单靶点和多靶点基因组编辑。