Griffith Elen
MRC Human Genetics Unit, Western General Hospital, Edinburgh, UK.
Methods Mol Biol. 2007;370:97-108. doi: 10.1007/978-1-59745-353-0_8.
RNA interference (RNAi) is a specific and efficient method to knock down protein levels using small interfering RNAs (siRNAs), which target mRNA degradation. RNAi can be used in mammalian cell culture systems to target any protein of interest, and several studies have used this method to knock down adhesion proteins. We used siRNAs to knock down the levels of TES, a focal adhesion protein, in HeLa cells. We demonstrated knockdown of both TES mRNA and TES protein. Although total knockdown of TES was not achieved, the observed reduction in TES protein was sufficient to result in a cellular phenotype of reduced actin stress fibers.
RNA干扰(RNAi)是一种利用小干扰RNA(siRNA)降低蛋白质水平的特异性高效方法,siRNA靶向mRNA降解。RNAi可用于哺乳动物细胞培养系统,以靶向任何感兴趣的蛋白质,并且有多项研究使用该方法降低黏附蛋白水平。我们使用siRNA在HeLa细胞中降低黏着斑蛋白TES的水平。我们证明了TES mRNA和TES蛋白水平均被降低。虽然未实现TES的完全敲低,但观察到的TES蛋白减少足以导致肌动蛋白应力纤维减少的细胞表型。