Specificity and functionality of microRNA inhibitors.
Specificity and functionality of microRNA inhibitors.
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DOI：
10.1186/1758-907x-1-10
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发表时间：
2010-04-01
期刊：
影响因子：
--
通讯作者：
Vermeulen A
中科院分区：
文献类型：
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作者：
Robertson B;Dalby AB;Karpilow J;Khvorova A;Leake D;Vermeulen A
微RNA （mi）通过翻译衰减和信使RNA (m)降解调节基因表达，并与分化、体内平衡和疾病有关。天然miRNA靶标识别主要由种子区（核苷酸位置2至7）的完美互补决定，其他相互作用以序列和靶标特异性的方式起作用。合成的miRNA靶类似物是完全互补的，化学修饰的寡核苷酸，已经成功地用于抑制miRNA的功能。在本文中，我们提出了第一个系统的研究，以评估靶位错配对合成抑制剂活性的影响。针对三种miRNA （miR-21， miR-22和miR-122）测试了含有两核苷酸错配的miRNA抑制剂组。结果表明，抑制剂的功能随其失配位置的变化而变化。数据表明，天然miRNA目标识别的重要特征（如种子区互补性）对抑制剂的功能也很重要。此外，在第二，更多的3'区域的碱基配对似乎在决定合成抑制剂的功效方面同样重要。考虑到这些抑制区域的重要性和密切相关的miRNA序列的表达将使研究人员在未来的实验中更准确地解释结果。
Micro(mi)RNAs regulate gene expression through translational attenuation and messenger (m)RNA degradation, and are associated with differentiation, homeostasis and disease. Natural miRNA target recognition is determined primarily by perfect complementarity in a seed region (nucleotide positions 2 to 7) with additional interactions contributing in a sequence- and target-specific manner. Synthetic miRNA target analogs, which are fully complementary, chemically modified oligonucleotides, have been used successfully to inhibit miRNA function. In this paper, we present a first systematic study to evaluate the effect of mismatches in the target site on synthetic inhibitor activity. Panels of miRNA inhibitors containing two-nucleotide mismatches across the target site were tested against three miRNAs (miR-21, miR-22 and miR-122). The results showed that the function of inhibitors vary as mismatch positions in the inhibitors change. The data indicate that features important for natural miRNA target recognition (such as seed region complementarity) are also important for inhibitor functionality. In addition, base pairing at a second, more 3' region appears to be equally important in determining the efficacy of synthetic inhibitors. Considering the importance of these inhibitor regions and the expression of closely related miRNA sequences will enable researchers to interpret results more accurately in future experiments.