Silencing of microRNA families by seed-targeting tiny LNAs.
Silencing of microRNA families by seed-targeting tiny LNAs.
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理解动物microRNA（miRNAs）的广泛生物学作用的挑战促进了用于miRNAs功能丧失研究的遗传和功能基因组学技术的发展。然而，用于探索整个miRNA家族功能的工具仍然有限。我们开发了一种使用种子靶向8-mer锁核酸（LNA）寡核苷酸（称为微小LNA）拮抗miRNA功能的方法。将微小LNA转染到细胞中导致共享相同种子的家族内的miRNA的同时抑制，伴随着直接靶标的上调。此外，全身递送的未缀合的微小LNA在小鼠的许多正常组织和乳腺肿瘤中显示出摄取，与长期miRNA沉默一致。转录和蛋白质组学分析表明，微小LNA具有可忽略的脱靶效应，不会显著改变具有完美微小LNA互补位点的mRNA的输出。综合考虑，这些数据支持微小LNA在阐明体内miRNA家族功能中的效用。
The challenge of understanding the widespread biological roles of animal microRNAs (miRNAs) has prompted the development of genetic and functional genomics technologies for miRNA loss-of-function studies. However, tools for exploring the functions of entire miRNA families are still limited. We developed a method that enables antagonism of miRNA function using seed-targeting 8-mer locked nucleic acid (LNA) oligonucleotides, termed tiny LNAs. Transfection of tiny LNAs into cells resulted in simultaneous inhibition of miRNAs within families sharing the same seed with concomitant upregulation of direct targets. In addition, systemically delivered, unconjugated tiny LNAs showed uptake in many normal tissues and in breast tumors in mice, coinciding with long-term miRNA silencing. Transcriptional and proteomic profiling suggested that tiny LNAs have negligible off-target effects, not significantly altering the output from mRNAs with perfect tiny LNA complementary sites. Considered together, these data support the utility of tiny LNAs in elucidating the functions of miRNA families in vivo.