用CRISPR/Cas9系统构建d XPR1的果蝇突变体品系  

Generate a dXPR1 null fly stock with the CRISPR/Cas9 system

作　　者：王睿龙 牛晓晓[1] 金珊[1] WANG Ruilong;NIU Xiaoxiao;JIN Shan(College of Life Sciences,Hubei University,Wuhan 430062,China)

机构地区：[1]湖北大学生命科学学院

出　　处：《湖北大学学报（自然科学版）》2020年第1期27-31,共5页Journal of Hubei University：Natural Science

基　　金：国家自然科学基金(81871121)资助

摘　　要：XPR 1是一种细胞无机磷酸盐输出蛋白的编码基因,其突变会导致原发型家族性脑钙化(primary familial brain calcification,PFBC).XPR 1在果蝇中同源基因为CG10483(dXPR1).使用CRISPR/Cas9系统,通过构建引导Cas9内切酶的sgRNA表达载体,并将其注射到表达Cas9酶的果蝇卵中,构建了dXPR 1大片段缺失果蝇品系即dXPR 1无功能突变体(dXPR 1^-).通过观察dXPR 1^-的表型从而初步判断XPR 1基因对神经系统发育的影响,为进一步的XPR 1基因功能与致病机制的研究提供基础.Human XPR 1 is an inorganic phosphate export protein encoding gene.The mutations of XPR 1 can result in primary familial brain calcification(PFBC).The homologous of human XPR 1 is CG 10483(dXPR 1)in Drosophila.In our researches,the CRISPR/Cas9 system was used to generate a large fragment-deficiency mutant in dXPR 1,dXPR 1^-mutant,in Drosophila.SgRNAs that direct Cas9 endonuclease to edit the XPR 1 target sequence,were constructed into vectors and injected into fly eggs,which expressed Cas9 enzyme.The phenotype of dXPR 1^-was observed to preliminarily determine the influence of XPR 1 gene on the development of the nervous system,providing a basis for further researches on the function and pathogenesis of XPR 1 gene.

关 键 词：果蝇 PFBC CRISPR/Cas9系统 XPR1 dXPR1^- 

分 类 号：Q789[生物学—分子生物学]