Inhibition of RNAse A family enzymes prevents degradation and loss of silencing activity of siRNAs in serum
Inhibition of RNAse A family enzymes prevents degradation and loss of silencing activity of siRNAs in serum
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DOI：
10.1016/j.bcp.2005.11.015
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发表时间：
2006-02-28
影响因子：
5.8
通讯作者：
Piiper, A
中科院分区：
文献类型：
--
作者：
Haupenthal, J;Baehr, C;Piiper, A
小干扰RNA（siRNA），类似于21个核苷酸的RNA双链体，通过称为RNA干扰的过程提供了一种有前途的方法来特异性地降解靶细胞中的RNA。体内稳定性不足是siRNA在人体中系统应用的主要问题。本研究表明，RNA酶A样RNA酶降解血清中的siRNA。siRNA对血清中降解的敏感性通过siRNA序列内A/U的局部聚类（即显示低热稳定性的区域，最显著地在分子的末端）和T-突出碱基而强烈增强。重要的是，RNA酶A家族酶的抑制防止了siRNA在血清中的降解和沉默活性的丧失。此外，siRNA在人血清中的降解比在小鼠血清中的降解快得多，这表明RNA酶A家族酶对siRNA的降解可能是siRNA在人中的未来治疗应用中比在小鼠模型中更具挑战性的问题。总之，本研究表明siRNA在血清中被RNA酶A家族酶降解，并且它们在血清中降解的动力学取决于它们的序列。这些发现可能对siRNA未来可能的人类治疗应用非常重要。(c)2005年爱思唯尔公司All rights reserved.
Small interfering RNAs (siRNA), RNA duplexes of similar to 21 nucleotides, offer a promising approach to specifically degrade RNAs in target cells by a process termed RNA interference. Insufficient in vivo-stability is a major problem of a systemic application of siRNAs in humans. The present study demonstrated that RNAse A-like RNAses degraded siRNAs in serum. The susceptibility of siRNAs towards degradation in serum was strongly enhanced by local clustering of A/Us within the siRNA sequence, i.e. regions showing low thermal stability, most notably at the ends of the molecule, and by T-overhanging bases. Importantly, inhibition of RNAse A family enzymes prevented the degradation and loss of silencing activity of siRNAs in serum. Furthermore, the degradation of siRNAs was considerably faster in human than in mouse serum, suggesting that the degradation of siRNAs by RNAse A family enzymes might be a more challenging problem in a future therapeutic application of siRNAs in humans than in mouse models. Together, the present study indicates that siRNAs are degraded by RNAse A family enzymes in serum and that the kinetics of their degradation in serum depends on their sequence. These findings might be of great importance for a possible future human therapeutic application of siRNAs. (c) 2005 Elsevier Inc. All rights reserved.