Login
欢迎浏览恩派尔生物资料网
我要投稿 请登录 免费注册 安全退出

您现在的位置是: 首页 > 实验方法 > 生物化学

生物化学

A rapid and sensitive assay for quantification of siRNA efficiency and specificity

2025-03-27 生物化学 加入收藏
RNA Interference has rapidly emerged as an efficient procedure for knocking down

RNA Interference has rapidly emerged as an efficient procedure for knocking down gene expression in model systems. However, cross-reactivity, whereby multiple genes may be simultaneously targeted by a single short interfering RNA (siRNA), can potentially jeopardize correct interpretation of gene function. As such, it is essential to test the specificity of a siRNA prior to a full phenotypic analysis. To this end, we have adapted a reporter-based assay harnessing the sensitivity of luciferase activity to provide a quantitative readout of relative RNAi efficacy and specificity. We have tested different siRNAs directed against Thymosin β4 (Tβ4 ); determined their effectiveness at silencing Tβ4 and have both excluded off-target silencing of the Tβ4 homologue Thymosin β10 (Tβ10 ) and demonstrated partial knockdown of Tβ10 despite significant (12/23; 52%) sequence mismatch. This assay system is applicable to any RNAi study where there is a risk of targeting homologous genes and to the monitoring of off-target effects at the genome level following microarray expression profiling.

文章底部广告位

文章评论

加载中~