Innovative Biology Laboratories, Neuroscience Drug Discovery Unit, Research, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan; Shonan Incubation Laboratories, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan.
Drug Safety Research Laboratories, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan.
J Biol Chem. 2021 Jan-Jun;296:100409. doi: 10.1016/j.jbc.2021.100409. Epub 2021 Feb 10.
Microprocessor complex, including DiGeorge syndrome critical region gene 8 (DGCR8) and DROSHA, recognizes and cleaves primary transcripts of microRNAs (pri-miRNAs) in the maturation of canonical miRNAs. The study of DGCR8 haploinsufficiency reveals that the efficiency of this activity varies for different miRNA species. It is thought that this variation might be associated with the risk of schizophrenia with 22q11 deletion syndrome caused by disruption of the DGCR8 gene. However, the underlying mechanism for varying action of DGCR8 with each miRNA remains largely unknown. Here, we used in vivo monitoring to measure the efficiency of DGCR8-dependent microprocessor activity in cultured cells. We confirmed that this system recapitulates the microprocessor activity of endogenous pri-miRNA with expression of a ratiometric fluorescence reporter. Using this system, we detected mir-9-2 as one of the most efficient targets. We also identified a novel DGCR8-responsive RNA element, which is highly conserved among mammalian species and could be regulated at the epi-transcriptome (RNA modification) level. This unique feature between DGCR8 and pri-miR-9-2 processing may suggest a link to the risk of schizophrenia.
微处理器复合物，包括 DiGeorge 综合征关键区基因 8（DGCR8）和 DROSHA，在成熟的典型 miRNA 中识别和切割 microRNAs（pri-miRNAs）的初级转录物。对 DGCR8 杂合不足的研究表明，这种活性对于不同的 miRNA 物种的效率不同。人们认为，这种差异可能与 DGCR8 基因破坏引起的 22q11 缺失综合征导致精神分裂症的风险有关。然而，DGCR8 与每个 miRNA 不同作用的潜在机制在很大程度上仍然未知。在这里，我们使用体内监测来测量培养细胞中 DGCR8 依赖性微处理器活性的效率。我们证实，该系统通过表达比率荧光报告基因来重现内源性 pri-miRNA 的微处理器活性。使用该系统，我们检测到 mir-9-2 是最有效的靶标之一。我们还鉴定了一种新的 DGCR8 反应性 RNA 元件，该元件在哺乳动物物种中高度保守，并且可以在 epi-transcriptome（RNA 修饰）水平上进行调控。DGCR8 和 pri-miR-9-2 加工之间的这种独特特征可能表明与精神分裂症的风险有关。
