摘要
畜牧业是农业的重要组成部分,其经济价值与畜禽表型性状(如产肉量、产蛋率等)密切相关。这些性状由复杂的遗传网络调控,并且受到环境因素的影响,导致挖掘决定这些性状的功能基因较为困难。遗传筛选技术的出现使基因型-表型关系的发现变得简单,但早期的遗传筛选技术通量低并且难以分析产生的大量数据致使效率低下。随着生物技术的迭代更新,遗传筛选技术进入高通量筛选时代,为解析基因型-表型关系提供了高效手段。传统的高通量筛选方法主要包括RNA干扰(RNAi)和cDNA过表达等,但RNAi易脱靶且无法完全抑制基因表达,cDNA文库构建成本高且可能引发细胞毒性。CRISPR/Cas系统的开发促使为遗传筛选提供了新的策略。该系统基于细菌的适应性免疫机制,通过向导RNA靶向DNA序列并利用Cas蛋白精准编辑基因,具有操作简便、特异性高和适用范围广的优势。CRISPR高通量筛选技术主要包括功能缺失型(CRISPRko)、功能获得型(CRISPRa/CRISPRi)、碱基编辑等多种模式,能够直接在DNA水平进行扰动,克服了传统方法的不足。本文介绍了遗传筛选技术的类型、特点以及在畜牧学中的应用,并展望了CRISPR高通量筛选技术在畜牧业育种中的巨大潜力。
Animal husbandry is an important part of agriculture,and its economic value is closely related to the phenotypic traits of livestock and poultry(such as meat production,egg production rate,etc.).These traits are regulated by complex genetic networks and are affected by environmental factors,causing it difficult to mine functional genes that determine these traits.The emergence of genetic screening technology makes the discovery of genotype-phenotype relationship simple,but the early genetic screening technology has low throughput and is difficult to analyze the large amount of data generated,resulting in low efficiency.With the iterative update of biotechnology,genetic screening technology has entered the era of high-throughput screening,which provides an efficient means for analyzing genotype-phenotype relationship.Traditional high-throughput screening methods mainly include RNA interference(RNAi)and cDNA overexpression.However,RNAi is prone to have off-target and cannot completely inhibit gene expression.The construction of cDNA library is costly and may cause cytotoxicity.The development of CRISPR/Cas system provides a new strategy for genetic screening.Based on the adaptive immune mechanism of bacteria,the system targets DNA sequences by guide RNA and accurately edits genes by Cas protein,which has the advantages of simple operation,high specificity and wide application range.CRISPR high-throughput screening technology mainly includes multiple modes such as loss-of-function(CRISPRko),gain-of-function(CRISPRa/CRISPRi),and base editing,which can directly perturb at the DNA level and overcome the shortcomings of traditional methods.This paper briefly introduces the types and characteristics of genetic screening technology and its application in animal husbandry,and looks forward to the great potential of CRISPR high-throughput screening technology in animal husbandry breeding.
作者
田云
孔辰
杨冲
刘统高
高红瑞
李佳慧
马云
蔡蓓
TIAN Yun;KONG Chen;YANG Chong;LIU Tong-gao;GAO Hong-rui;LI Jia-hui;MA Yun;CAI Bei(Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region,College of Animal Science and Technology,Ningxia University,Yinchuan 750021;Ningxia Hui Autonomous Region Animal Husbandry Workstation,Yinchuan 750002)
出处
《生物技术通报》
北大核心
2025年第12期82-94,共13页
Biotechnology Bulletin
基金
国家自然科学基金项目(022304370026)
宁夏重点研发项目(022204000135,2023BCF01005)。
