利用载体介导小干扰RNA诱导RSRC1基因沉默

目的：设计并构建人RSRC1基因小干扰RNA（siRNA）的真核表达载体,并观察其沉默效果。方法：以人RSRC1基因的cDNA序列为靶标,设计含有小发卡结构的2条寡核苷酸序列,并将其克隆到siRNA表达载体pSliencer2.1-U6neo上,转化大肠杆菌DH5α菌株,抽提质粒,测序正确后将重组质粒转染人胚肾293T细胞,通过Western blot和荧光分析检测其抑制效果。结果：重组体测序成功后,Western blot分析证明构建的siRNA能有效抑制外源性及内源性RSRC1表达;将siRNA重组质粒和带GFP标签的RSRC1共转染293T细胞,荧光显微镜下GFP的亮度明显减弱。结论：获得了2条人RSRC1siRNA真核表达载体,均能有效地抑制RSRC1基因表达。

Inhibition of the Expression of the Human Arginine/Serine-Rich Coiled-Coil 1 Gene by Small Interference RNA

Objective： To design and construct the small interference RNA（siRNA） for RSRC1 gene and investigate the interference effect on the expression of RSRC1. Methods： siRNAs were designed and constructed based on human RSRC1 cDNA sequence. Two pairs of oligonucleotides were designed and inserted into pSlieneer 2.1-U6 neo expression vector. After confirmation of the constructs by restriction digestion and DNA sequencing, human 293T cells were eo-transfected with the recombination plasmids and FLAG-tagged RSRC1 expression vector. The dumbness effect of RSRC1 siRNAs on the expression of RSRC1 was identified by Western blot and fluorescence intensity. Results： The two RSRC1 siRNAs were obtained and could effectively inhibit the expression of RSRC1. Conclusion： The two constructed RSRC1 siRNAs could ef- fectively inhibit RSRC1 expression.