生物技术通报2025，Vol.41Issue(12)：82-94,13.DOI:10.13560/j.cnki.biotech.bull.1985.2025-0500
CRISPR高通量筛选技术及其在畜牧研究中的应用
CRISPR-based High-throughput Screening Technology and Its Applications in Livestock Research
摘要
Abstract
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.关键词
畜牧业/遗传筛选/CRISPR高通量筛选/SgRNA文库Key words
animal husbandry/genetic screening/CRISPR high-throughput screening/SgRNA library引用本文复制引用
TIAN Yun,KONG Chen,YANG Chong,LIU Tong-gao,GAO Hong-rui,LI Jia-hui,MA Yun,CAI Bei..CRISPR高通量筛选技术及其在畜牧研究中的应用[J].生物技术通报,2025,41(12):82-94,13.基金项目
国家自然科学基金项目(022304370026),宁夏重点研发项目(022204000135,2023BCF01005) （022304370026）
