CRISPR/Cas9 knock-in介导的输卵管特异性表达转基因鸡研究进展  

Progress on CRISPR/Cas9 Knock-in Mediated Transgenic Chickens with Oviduct-specific Expression

作　　者：刘灵康 郑喜邦[1] LIU Lingkang;ZHENG Xibang(College of Animal Science and Technology,Guangxi University,Nanning,Guangxi 530004)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004

出　　处：《中国家禽》2020年第3期75-81,共7页China Poultry

基　　金：广西自然科学基金(2018GXNSFDA281026);国家自然科学基金(31660653);广西科技计划重点研发项目(桂科AB16380098)。

摘　　要：利用动物生物反应器生产重组蛋白是一种具有应用前景的生物技术。鸡输卵管生物反应器是理想的动物生物反应器之一,其优点在于表达的外源蛋白能够分泌到蛋清中,可避免蛋白提取过程中对鸡本身造成伤害,同时蛋清成分简单,便于后期的纯化。目前利用慢病毒结合原始生殖细胞(PGCs)制备转基因鸡被认为是最可行的方法,但因外源基因随机整合且生殖系传递效率较低,使转基因鸡研究受到技术上的限制。而2013年问世的CRISPR/Cas9基因敲入(CRISPR/Cas9 knock-in)技术能够使外源基因精准定向插入基因组特异性位点,这对生产输卵管特异性转基因鸡具有重大意义。文章综述了鸡输卵管反应器的研究进展、CRISPR/Cas9 knock-in技术在输卵管特异性表达转基因鸡研究和鸡育种领域的应用现状,并指出了目前存在的问题和相应的解决办法。Producing recombinant proteins through animal bioreactors is a promising technology.Chicken oviduct bioreactor is one of the ideal animal bioreactors.Its advantages include:the interest of protein(IOP)expressed in oviduct and deposited in egg white,and no harm is done to chicken itself during the extraction and purification of the targeting proteins;the composition of the egg white is relatively simple,and the IOP purification is done easily.To date,combination of lentivirus and primordial germ cells(PGCs)is considered to be a feasible method to generate transgenic chickens.Due to the random integration of exogenous genes to host genome,and the lower germline transmission efficiency of transgenes caused by lentiviral vectors,production of transgenic chicken is limited technologically.Nevertheless,the appearance of CRISPR/Cas9 knock-in technology in 2013 overcomes the problems.It integrates transgenes precisely to the desired locus of host genome,and is of great significance for the production of transgenic chickens with oviduct-specific expression.This paper reviews the progress of chicken oviduct bioreactor,the application of CRISPR/Cas9 knock-in technology both in the field of transgenic chickens with oviduct-specific expression,and in that of chicken breeding industry.Finally,the difficulties in transgenic chicken are discussed,and suggestions are given to fight them.

关 键 词：转基因鸡 原始生殖细胞 CRISPR/Cas9 基因敲入 输卵管特异性表达 

分 类 号：S831[农业科学—畜牧学] Q78[农业科学—畜牧兽医]