RNA interference (RNAi) has great potential as a tool for studying gene function in mammals. However, the specificity and magnitude of the in vivo response to RNAi remains to be fully characterized. A molecular and phenotypic comparison of a genetic knockout mouse and the corresponding knockdown version would help clarify the utility of the RNAi approach. Here, we used hydrodynamic delivery of small interfering RNA (siRNA) to knockdown peroxisome proliferator activated receptor alpha (Ppara), a gene that is central to the regulation of fatty acid metabolism. We found that Ppara knockdown in the liver results in a transcript profile and metabolic phenotype that is comparable to those of Ppara-/- mice. Combining the profiles from mice treated with the PPARalpha agonist fenofibrate, we confirmed the specificity of the RNAi response and identified candidate genes proximal to PPARalpha regulation. Ppara knockdown animals developed hypoglycemia and hypertriglyceridemia, phenotypes observed in Ppara-/- mice. In contrast to Ppara-/- mice, fasting was not required to uncover these phenotypes. Together, these data validate the utility of the RNAi approach and suggest that siRNA can be used as a complement to classical knockout technology in gene function studies.

译文

RNA干扰(RNAi)作为研究哺乳动物基因功能的工具具有巨大潜力。但是,体内对RNAi应答的特异性和强度仍有待充分表征。基因敲除小鼠和相应的敲除版本的分子和表型比较将有助于阐明RNAi方法的实用性。在这里,我们利用小分子干扰RNA(siRNA)的流体动力学传递来敲低过氧化物酶体增殖物激活的受体α(Ppara),该基因是调节脂肪酸代谢的关键基因。我们发现肝脏中的Ppara敲低导致与Ppara-/-小鼠的转录谱和代谢表型相当。结合使用PPARalpha激动剂非诺贝特治疗的小鼠的概况,我们确认了RNAi反应的特异性,并确定了与PPARalpha调节最接近的候选基因。 Ppara基因敲低动物出现低血糖和高甘油三酯血症,这是在Ppara-/-小鼠中观察到的表型。与Ppara-/-小鼠相反,不需要禁食即可发现这些表型。总之,这些数据验证了RNAi方法的实用性,并表明siRNA可以用作基因功能研究中经典敲除技术的补充。

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