通过 CRISPR/Cas9 介导的 DNA 双链断裂在水稻中的基因靶向。
Gene Targeting via CRISPR/Cas9-Mediated DNA Double-Strand Break Induction in Rice.
机构信息
Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.
Graduate School of Nanobioscience, Yokohama City University, Kanagawa, Japan.
出版信息
Methods Mol Biol. 2025;2869:91-100. doi: 10.1007/978-1-0716-4204-7_10.
Gene targeting (GT) is a precise genome editing tool to achieve desired modification of a target gene, e.g., introduction of point mutations, knock-in of a reporter gene, or swapping of a functional domain, through homologous recombination. However, due to its low frequency, it has proved difficult to establish a universal GT system. The availability of the CRISPR/Cas9 system for genome editing in plants has opened up possibilities to apply GT successfully to some plant species. Here, we provide protocols for CRISPR/Cas9-mediated DNA double-strand break (DSB)-induced GT in rice.
基因打靶（GT）是一种精确的基因组编辑工具，可通过同源重组实现对靶基因的期望修饰，例如引入点突变、敲入报告基因或交换功能域。然而，由于其频率较低，因此建立通用的 GT 系统一直很困难。CRISPR/Cas9 系统在植物基因组编辑中的应用为 GT 在一些植物物种中的成功应用开辟了可能性。在这里，我们提供了用于水稻中 CRISPR/Cas9 介导的 DNA 双链断裂（DSB）诱导 GT 的方案。
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