miR-122 targeting with LNA/2′-O-methyl oligonucleotide mixmers, peptide nucleic acids (PNA), and PNA-peptide conjugates
miR-122 targeting with LNA/2′-O-methyl oligonucleotide mixmers, peptide nucleic acids (PNA), and PNA-peptide conjugates
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DOI：
10.1261/rna.844108
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发表时间：
2008-02-01
期刊：
影响因子：
4.5
通讯作者：
Gait, Michael J.
中科院分区：
文献类型：
--
作者：
Fabani, Martin M.;Gait, Michael J.
MicroRNAs是一种小的非编码RNA，以转录后模式调节许多细胞过程。用反义寡核苷酸敲除microRNA是探索microRNA功能和作为潜在治疗手段的有用策略。MicroRNA-122(miR-122)是一种肝脏特异的microRNA，其主要功能与脂质代谢和肝脏动态平衡有关。在这里，我们证明了基于锁定核酸(LNA)/2‘-O-甲基混合物的反义寡核苷酸或多肽核酸(PNA)寡聚体的电穿孔能够有效地阻断人和大鼠肝细胞中miR-122的活性。这些寡核苷酸类似物首次在抑制microRNA方面进行了评估，在结合和抑制microRNA作用方面比标准的2‘-O-甲基寡核苷酸更有效。我们还表明，无需转染或电穿孔人或大鼠细胞系，通过将反义PNA与细胞穿透肽R-6-穿透蛋白偶联，或仅通过与四个赖氨酸残基连接，就可以实现对microRNA的抑制，这突出了PNA在未来治疗应用以及研究microRNA功能方面的潜力。
MicroRNAs are small noncoding RNAs that regulate many cellular processes in a post-transcriptional mode. MicroRNA knockdown by antisense oligonucleotides is a useful strategy to explore microRNA functionality and as potential therapeutics. MicroRNA-122 (miR-122) is a liver-specific microRNA, the main function of which has been linked with lipid metabolism and liver homeostasis. Here, we show that lipofection of an antisense oligonucleotide based on a Locked Nucleic Acids (LNA)/2'-O-methyl mixmer or electroporation of a Peptide Nucleic Acid (PNA) oligomer is effective at blocking miR-122 activity in human and rat liver cells. These oligonucleotide analogs, evaluated for the first time in microRNA inhibition, are more effective than standard 2'-O-methyl oligonucleotides in binding and inhibiting microRNA action. We also show that microRNA inhibition can be achieved without the need for transfection or electroporation of the human or rat cell lines, by conjugation of an antisense PNA to the cell-penetrating peptide R-6-Penetratin, or merely by linkage to just four Lys residues, highlighting the potential of PNA for future therapeutic applications as well as for studying microRNA function.