讨教一个SiRNA问题

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请问哪位高手能够指点一二?
SiRNA干扰实验的模型通常采用GFP,采用GFP表达细胞株,设计SiRNA干扰序列Duplex,采用合适的转染试剂将SiRNA导入细胞,通过荧光显微镜观察GFP的表达,检测干扰效果。但是,由于GFP蛋白在细胞内的半衰期通常较长,所以采用该系统考察瞬时的干扰效果,不是很理想。请问是不是这样?如果是,请问还有那些报告基因的目的蛋白在细胞内的半衰期较短,适合作为SiRNA干扰研究的模型。

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most of the cell cylce proteins have rather short half life. look at some of the earlier publications where the 'power' of siRNA was first described in mammalian cells, i think you can use any of the proteins they used in the paper.

blu 的头像

如果指标蛋白半衰期长的话, SiRNA的抑制效果就不容易测到. 在加入SiRNA之前细胞内是有指标蛋白表达的, 而SiRNA的抑制效果是短时性的(即时转染). 在SiRNA转染前细胞已经表达的蛋白如果没有降解彻底,抑制的效果就不明显了.