CRISPR/Cas：基因功能研究和作物改良的有力工具。

CRISPR/Cas: A powerful tool for gene function study and crop improvement.

机构信息

Henan Province Engineering Research Center for Forest Biomass Value-Added Products, College of Forestry, Henan Agricultural University, Zhengzhou, Henan 450002, China.

Henan Collaborative Innovation Center of Modern Biological Breeding and Henan Key Laboratory for Molecular Ecology and Germplasm Innovation of Cotton and Wheat, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China.

出版信息

J Adv Res. 2020 Oct 21;29:207-221. doi: 10.1016/j.jare.2020.10.003. eCollection 2021 Mar.

DOI:10.1016/j.jare.2020.10.003

PMID:33842017

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8020163/

Abstract

BACKGROUND

It is a long-standing goal of scientists and breeders to precisely control a gene for studying its function as well as improving crop yield, quality, and tolerance to various environmental stresses. The discovery and modification of CRISPR/Cas system, a nature-occurred gene editing tool, opens an era for studying gene function and precision crop breeding.

AIM OF REVIEW

In this review, we first introduce the brief history of CRISPR/Cas discovery followed the mechanism and application of CRISPR/Cas system on gene function study and crop improvement. Currently, CRISPR/Cas genome editing has been becoming a mature cutting-edge biotechnological tool for crop improvement that already used in many different traits in crops, including pathogen resistance, abiotic tolerance, plant development and morphology and even secondary metabolism and fiber development. Finally, we point out the major issues associating with CRISPR/Cas system and the future research directions. CRISPR/Cas9 system is a robust and powerful biotechnological tool for targeting an individual DNA and RNA sequence in the genome. It can be used to target a sequence for gene knockin, knockout and replacement as well as monitoring and regulating gene expression at the genome and epigenome levels by binding a specific sequence. -mediated method is still the major and efficient method for delivering CRISPR/Cas regents into targeted plant cells. However, other delivery methods, such as virus-mediated method, have been developed and enhanced the application potentials of CRISPR/Cas9-based crop improvement. PAM requirement offers the CRISPR/Cas9-targted genetic loci and also limits the application of CRISPR/Cas9. Discovering new Cas proteins and modifying current Cas enzymes play an important role in CRISPR/Cas9-based genome editing. Developing a better CRISPR/Cas9 system, including the delivery system and the methods eliminating off-target effects, and finding key/master genes for controlling crop growth and development is two major directions for CRISPR/Cas9-based crop improvement.

摘要

背景

长期以来，科学家和育种家的目标是精确地控制一个基因，以研究其功能，并提高作物的产量、质量和对各种环境胁迫的耐受性。CRISPR/Cas 系统的发现和修饰，一种天然存在的基因编辑工具，开启了研究基因功能和精准作物育种的时代。

综述目的

在这篇综述中，我们首先介绍了 CRISPR/Cas 的简要发现历史，以及 CRISPR/Cas 系统在基因功能研究和作物改良中的机制和应用。目前，CRISPR/Cas 基因组编辑已经成为一种成熟的前沿生物技术工具，用于作物改良，已经在许多不同的作物性状中得到应用，包括抗病性、非生物胁迫耐受性、植物发育和形态，甚至次生代谢和纤维发育。最后，我们指出了与 CRISPR/Cas 系统相关的主要问题和未来的研究方向。CRISPR/Cas9 系统是一种强大的生物技术工具，可用于靶向基因组中的单个 DNA 和 RNA 序列。它可用于靶向特定序列进行基因敲入、敲除和替换，以及通过结合特定序列在基因组和表观基因组水平上监测和调节基因表达。基于 sgRNA 的方法仍然是将 CRISPR/Cas 试剂递送到靶标植物细胞的主要和有效方法。然而，已经开发并增强了基于 CRISPR/Cas9 的作物改良应用潜力的其他递送方法，如病毒介导的方法。PAM 要求提供了 CRISPR/Cas9 靶向的遗传基因座，也限制了 CRISPR/Cas9 的应用。发现新的 Cas 蛋白和修饰当前的 Cas 酶在基于 CRISPR/Cas9 的基因组编辑中发挥着重要作用。开发更好的 CRISPR/Cas9 系统，包括递送系统和消除脱靶效应的方法，以及寻找控制作物生长和发育的关键/主基因，是基于 CRISPR/Cas9 的作物改良的两个主要方向。