I 型 CRISPR 系统介导的植物基因组编辑。

Type I-D CRISPR System-Mediated Genome Editing in Plants.

机构信息

Graduate School of Technology, Industrial and Social Sciences, Tokushima University, Tokushima, Japan.

School of Life Science and Technology, Tokyo Institute of Technology, Kanagawa, Japan.

出版信息

Methods Mol Biol. 2023;2653:21-38. doi: 10.1007/978-1-0716-3131-7_2.

DOI:10.1007/978-1-0716-3131-7_2

PMID:36995617

Abstract

Genome editing has revolutionized plant research and plant breeding by enabling precise genome manipulation. In particular, the application of type II CRISPR-Cas9 systems to genome editing has proved an important milestone, accelerating genetic engineering and the analysis of gene function. On the other hand, the potential of other types of CRISPR-Cas systems, especially many of the most abundant type I CRISPR-Cas systems, remains unexplored. We recently developed a novel genome editing tool, TiD, based on the type I-D CRISPR-Cas system. In this chapter, we describe a protocol for genome editing of plant cells using TiD. This protocol allows the application of TiD to induce short insertion and deletions (indels) or long-range deletions at target sites with high specificity in tomato cells.

摘要

基因组编辑通过实现精确的基因组操作，彻底改变了植物研究和植物育种。特别是，II 型 CRISPR-Cas9 系统在基因组编辑中的应用被证明是一个重要的里程碑，加速了基因工程和基因功能分析。另一方面，其他类型的 CRISPR-Cas 系统的潜力，特别是许多最丰富的 I 型 CRISPR-Cas 系统，仍未得到探索。我们最近开发了一种基于 I 型-D CRISPR-Cas 系统的新型基因组编辑工具 TiD。在本章中，我们描述了使用 TiD 编辑植物细胞基因组的方案。该方案允许 TiD 在番茄细胞中靶向位点特异性诱导短插入和缺失（indels）或长片段缺失。