Vps28基因敲除小鼠模型的构建及其对泌乳和免疫性状影响的研究  

Construction of Vps28 Knock-out Mice and Model Study of the Impact on Lactation and Immune Traits

作　　者：丁亚群 丁宁 谢深民 黄梦娜 张昱 张勤 姜力 DING Ya-qun;DING Ning;XIE Shen-min;HUANG Meng-na;ZHANG Yu;ZHANG Qin;JIANG Li(National Engineering Laboratory for Animal Breeding,Key laboratory of Animal Genetics,Breeding and Reproduction,Ministry of Agriculture and Rural Affairs,College of Animal Science and Technology,China Agricultural University,Beijing 100193;College of Animal Science and Technology,Shandong Agricultural University,Tai’an 271018)

机构地区：[1]中国农业大学动物科学技术学院畜禽育种国家工程实验室农业农村部动物遗传育种与繁殖重点实验室,北京100193 [2]山东农业大学动物科技学院,泰安271018

出　　处：《生物技术通报》2022年第3期164-172,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31872327);北京市自然科学基金项目(6182023)。

摘　　要：尽管基于QTL定位和关联分析研究挖掘了许多奶牛产奶性状候选功能基因,然而仅有少数几个基因经过体内外的功能验证。本课题组前期全基因组关联分析(GWAS)研究结果表明Vps28基因与奶牛产奶性状表型显著相关,并在奶牛乳腺组织中特异性高表达。为了深入了解该基因对泌乳性状的作用和功能,本研究利用CRISPR/Cas9技术构建了Vps28基因敲除小鼠,对其进行体内功能研究。研究结果显示Vps28基因敲除纯合型小鼠(Vps28-/-)极可能导致胚胎早期死亡,故只获得杂合型基因敲除小鼠(Vps28+/-)。敲除小鼠模型构建成功后,检测乳腺组织和脾脏组织中Vps28基因mRNA和蛋白的表达,结果显示与野生型小鼠(Vps28+/+)相比,Vps28+/-杂合型小鼠Vps28的mRNA和蛋白表达量均显著下降。进一步对母鼠的泌乳量、乳房形态和血常规进行检测,发现Vps28敲除母鼠的泌乳量下降,乳头明显凹陷,且血常规中白细胞数量、中性粒细胞数量、淋巴细胞数量和中间细胞数量显著低于野生型小鼠,推测Vps28基因不但影响泌乳,而且影响相关免疫性状。Although many candidate genes for milk production traits have been mined based on QTL mapping and association analysis,only a few genes have been verified in vivo and in vitro.The results of our previous genome-wide association study(GWAS)showed that Vps28 was significantly associated with milk production traits in dairy cows.In addition,Vps28 was highly expressed in the mammary gland tissue of dairy cows.To further explore the role and function of Vps28 on milk production traits,CRISPR/Cas9 technology was used to construct Vps28 knockout mice,and its functions in vivo were studied.The results showed that Vps28 knockout homozygous mouse(Vps28-/-)were very likely to cause early embryonic lethality.Thus only heterozygous knock-out mice(Vps28+/-)were obtained in this study.Our results showed that the mRNA and protein expression in the mammary gland and spleen of Vps28 knockout mice(Vps28+/-)significantly decreased if compared with wild-type mice.Furthermore,Vps28 knockout mice had nipple dysplasia and reduced milk production.Also the white blood cells count,neutrophilic granulocyte count,lymphocyte count and intermediate cell count in the Vps28 knock-out mice were significantly lower than those in wild-type mice.These results suggest that Vps28 not only affects lactation,but also related immune traits in mice.

关 键 词：CRISPR/Cas9 基因敲除小鼠 Vps28基因 泌乳 免疫性状 

分 类 号：Q78[生物学—分子生物学] S823[农业科学—畜牧学]