CRISPR 介导的病毒基因敲除技术用于研究病毒逃避抗病毒先天免疫的机制。

CRISPR-Mediated Viral Gene Knockout to Investigate Viral Evasion of Antiviral Innate Immunity.

机构信息

Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Fujian Normal University, Fuzhou, China.

Department of Microbiology, Immunology & Infection Diseases, University of Calgary, Calgary, AB, Canada.

出版信息

Methods Mol Biol. 2025;2854:41-50. doi: 10.1007/978-1-0716-4108-8_6.

DOI:10.1007/978-1-0716-4108-8_6

PMID:39192117

Abstract

The innate immune system relies on a variety of pathogen recognition receptors (PRRs) as the first line of defense against pathogenic invasions. Viruses have evolved multiple strategies to evade the host immune system through coevolution with hosts. The CRISPR-Cas system is an adaptive immune system in bacteria or archaea that defends against viral reinvasion by targeting nucleic acids for cleavage. Based on the characteristics of Cas proteins and their variants, the CRISPR-Cas system has been developed into a versatile gene-editing tool capable of gene knockout or knock-in operations to achieve genetic variations in organisms. It is now widely used in the study of viral immune evasion mechanisms. This chapter will introduce the use of the CRISPR-Cas9 system for editing herpes simplex virus 1 (HSV-1) genes to explore the mechanisms by which HSV-1 evades host innate immunity and the experimental procedures involved.

摘要

先天免疫系统依赖多种病原体识别受体（PRRs）作为抵御病原体入侵的第一道防线。病毒通过与宿主的共同进化，进化出多种策略来逃避宿主免疫系统。CRISPR-Cas 系统是细菌或古菌中的一种适应性免疫系统，通过靶向核酸进行切割来防御病毒的再次入侵。基于 Cas 蛋白及其变体的特性，CRISPR-Cas 系统已被开发成为一种多功能的基因编辑工具，能够进行基因敲除或敲入操作，从而在生物体中实现基因变异。它现在被广泛用于研究病毒的免疫逃逸机制。本章将介绍使用 CRISPR-Cas9 系统编辑单纯疱疹病毒 1（HSV-1）基因，以探索 HSV-1 逃避宿主先天免疫的机制以及涉及的实验程序。