Scholarlink

<p>The core miRNA biogenesis enzyme DICER1 sustains recurrent mutations in cancer that compromise its RNase IIIb domain, which cleaves 5p arms of precursor microRNA hairpins. However, the lack of knock-in models has limited fuller understanding. Here, we generated <i>DICER1</i>-knockout and <i>DICER1</i><sup>S1344L</sup> (homozygous and hemizygous) human embryonic stem cells; the latter is a noncatalytic substitution in RNase IIIa that impairs RNase IIIb activity. <i>DICER1</i> knockouts lack canonical miRNAs, while S1344L induces two trends: ablation of miRNA-5p strands and selective changes in miRNA-3p strands. Curiously, we recognized directional upregulation of miRNA-3p passenger strands, indicating a broad strand switch. We used multiple in vitro assays to show 3p-arm-nicked pre-miRNAs preferentially load miRNA-3p species into Argonaute, compared to corresponding duplexes. Moreover, activity assays, RNA-sequencing data and Argonaute mRNA profiling confirm that these confer increased repression capacity. These data expand the molecular consequences of <i>DICER1</i> hotspot mutations in cancer.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Human DICER1 hotspot mutation induces both loss and gain of miRNA function

  • 期刊论文

  • 发表日期：

    2025年11月4日

查看全文
  • David Jee,
  • Seungjae Lee,
  • Dapeng Yang,
  • Robert Rickert,
  • Renfu Shang,
  • Danwei Huangfu,
  • Eric C. Lai

摘要

The core miRNA biogenesis enzyme DICER1 sustains recurrent mutations in cancer that compromise its RNase IIIb domain, which cleaves 5p arms of precursor microRNA hairpins. However, the lack of knock-in models has limited fuller understanding. Here, we generated DICER1-knockout and DICER1S1344L (homozygous and hemizygous) human embryonic stem cells; the latter is a noncatalytic substitution in RNase IIIa that impairs RNase IIIb activity. DICER1 knockouts lack canonical miRNAs, while S1344L induces two trends: ablation of miRNA-5p strands and selective changes in miRNA-3p strands. Curiously, we recognized directional upregulation of miRNA-3p passenger strands, indicating a broad strand switch. We used multiple in vitro assays to show 3p-arm-nicked pre-miRNAs preferentially load miRNA-3p species into Argonaute, compared to corresponding duplexes. Moreover, activity assays, RNA-sequencing data and Argonaute mRNA profiling confirm that these confer increased repression capacity. These data expand the molecular consequences of DICER1 hotspot mutations in cancer.