In vivo RNA structure analysis has become a powerful tool in molecular biology, largely due to the coupling of an increasingly diverse set of chemical approaches with high-throughput sequencing. This has resulted in a transition from single target to transcriptome-wide approaches. However, these methods require sequencing depths that preclude studying low abundance targets, which are not sufficiently captured in transcriptome-wide approaches. Here we present a ligation-free method to enrich for low abundance RNA sequences, which improves the diversity of molecules analyzed and results in improved analysis. In addition, this method is compatible with any choice of chemical adduct or read-out approach. We utilized this approach to study an autoregulated event in the pre-mRNA of the splicing factor, muscleblind-like splicing regulator 1 (MBNL1).

中文翻译：

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RNA 结构探测可表征活细胞中低丰度前 mRNA 上的 RNA-蛋白质相互作用


体内 RNA 结构分析已成为分子生物学中的强大工具，这主要是由于日益多样化的化学方法与高通量测序的结合。这导致了从单一目标到转录组范围方法的转变。然而，这些方法需要测序深度，这妨碍了研究低丰度靶标，而全转录组方法无法充分捕获低丰度靶标。在这里，我们提出了一种无需连接的方法来富集低丰度 RNA 序列，从而提高了分析分子的多样性并改进了分析。此外，该方法与任何化学加合物或读出方法的选择兼容。我们利用这种方法来研究剪接因子、肌盲样剪接调节因子 1 (MBNL1) 的前体 mRNA 中的自动调节事件。