Delivery of small interfering RNA with a synthetic collagen poly(Pro-Hyp-Gly) for gene silencing in vitro and in vivo
Delivery of small interfering RNA with a synthetic collagen poly(Pro-Hyp-Gly) for gene silencing in vitro and in vivo
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DOI：
10.1111/j.1440-169x.2010.01206.x
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发表时间：
2010-10-01
影响因子：
2.5
通讯作者：
Noji, Sumihare
中科院分区：
文献类型：
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作者：
Adachi, Taro;Kawakami, Emi;Noji, Sumihare
通过小干扰RNA（siRNA）沉默基因表达已成为许多动物遗传分析的有力工具。然而，siRNA的快速降解及其在体内作用的有限持续时间要求有效的递送技术。在这里，我们描述了与合成胶原蛋白聚（Pro-Hyp-Gly）（SYCOL）复合的siRNA对核酸酶具有抗性，并在体外和体内有效地转移到细胞中，从而允许体内长期基因沉默。我们发现SYCOL介导的靶向肌肉生长抑制素的siRNA在小鼠骨骼肌中的局部应用，编码骨骼肌生长的负调节因子，在应用后几周内引起肌肉质量的显著增加。此外，体内施用抗荧光素酶siRNA/SYCOL复合物部分降低了体内异种移植肿瘤中的荧光素酶表达。这些结果表明基于SYCOL的非病毒递送方法可以是通过RNAi在体内以及体外敲低基因表达的可靠的简单方法。
Silencing gene expression by small interfering RNAs (siRNAs) has become a powerful tool for the genetic analysis of many animals. However, the rapid degradation of siRNA and the limited duration of its action in vivo have called for an efficient delivery technology. Here, we describe that siRNA complexed with a synthetic collagen poly(Pro-Hyp-Gly) (SYCOL) is resistant to nucleases and is efficiently transferred into cells in vitro and in vivo, thereby allowing long-term gene silencing in vivo. We found that the SYCOL-mediated local application of siRNA targeting myostatin, coding a negative regulator of skeletal muscle growth, in mouse skeletal muscles, caused a marked increase in the muscle mass within a few weeks after application. Furthermore, in vivo administration of an anti-luciferase siRNA/SYCOL complex partially reduced luciferase expression in xenografted tumors in vivo. These results indicate a SYCOL-based non-viral delivery method could be a reliable simple approach to knockdown gene expression by RNAi in vivo as well as in vitro.