利用 CRISPR-Cas9 进行精确基因组编辑。
Precision Genome Editing with CRISPR-Cas9.
机构信息
Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan.
Plant Genomics and Molecular Breeding Laboratory, National Institute for Biotechnology and Genetic Engineering College, Pakistan Institute of Engineering and Applied Sciences (NIBGE-C, PIEAS), Faisalabad, Punjab, Pakistan.
出版信息
Methods Mol Biol. 2024;2788:355-372. doi: 10.1007/978-1-0716-3782-1_21.
The CRISPR/Cas9 system is a revolutionary technology for genome editing that allows for precise and efficient modifications of DNA sequences. The system is composed of two main components, the Cas9 enzyme and a guide RNA (gRNA). The gRNA is designed to specifically target a desired DNA sequence, while the Cas9 enzyme acts as molecular scissors to cut the DNA at that specific location. The cell then repairs the digested DNA, either through nonhomologous end joining (NHEJ) or homology-directed repair (HDR), resulting in either indels or precise modifications of DNA sequences with broad implications in biotechnology, agriculture, and medicine. This chapter provides a practical approach for utilizing CRISPR/Cas9 in precise genome editing, including identifying the target gene sequence, designing gRNA and protein (Cas9), and delivering the CRISPR components to target cells.
CRISPR/Cas9 系统是一种革命性的基因组编辑技术，可实现 DNA 序列的精确和高效修饰。该系统由两个主要组件组成，Cas9 酶和向导 RNA（gRNA）。gRNA 被设计为特异性靶向所需的 DNA 序列，而 Cas9 酶则充当分子剪刀，在该特定位置切割 DNA。然后，细胞通过非同源末端连接（NHEJ）或同源定向修复（HDR）修复消化的 DNA，导致 DNA 序列的插入缺失或精确修饰，这在生物技术、农业和医学中有广泛的应用。本章提供了一种实用的方法，用于利用 CRISPR/Cas9 进行精确的基因组编辑，包括确定靶基因序列、设计 gRNA 和蛋白质（Cas9）以及将 CRISPR 组件递送到靶细胞。
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