用CRISPR/Cas9系统构建原位表达d XPR1-GFP的果蝇品系

Generating d XPR1-GFP stock in situ expressed by CRISPR/Cas9 system in Drosophila

XPR1是最近鉴定出的一个原发性家族性脑钙化症(primary familial brian calcification,PFBC)的致病基因,它在果蝇体内的同源基因是CG10483(dXPR1).使用CRISPR/Cas9系统,构建原位表达dXPR1-GFP的果蝇品系,在CG10483终止密码子之前插入绿色荧光蛋白(green fluorescent protein,GFP)的编码序列,使果蝇表达dXPR1-GFP融合蛋白.该融合蛋白既可以被CG10483编码蛋白的抗体和GFP的抗体识别又能在激发光下发出绿色荧光,能为dXPR1编码蛋白提供准确的定位,从而为研究XPR1基因的功能提供方便.

XPR1 is a recent identified PFBC diseasecausing gene. It was first discovered as a receptor for xenotropic leukemia in mice. Further studies have found that it has the ability to transport inorganic phosphorus out of cells,and is the only gene known which has this function. The mutation of XPR1 gene causes the intracellular inorganic phosphorus transport disruption,which further results in the increase of inorganic phosphorus concentration in the cell,and the combination of phosphorous and calcium can form precipitate particles,resulting in a series of pathological processes. The homologous gene of human XPR1 in Drosophila is CG10483( dXPR1). The CRISPR/Cas9 system is used to generate endogenesious dXPR1-GFP fly in our researches. This fusion protein could be checked with GFP fluorescence directly or stained by GFP antibodies. It will provide a convenience tool for identified the location of XPR1 and further the study of the function of XPR1 gene.