The transition from dividing progenitors to postmitotic motor neurons (MNs) is orchestrated by a series of events, which are mainly studied at the transcriptional level by analyzing the activity of specific programming transcription factors. Here, we identify a post-transcriptional role of a MN-specific transcriptional unit (MN2) harboring a lncRNA (lncMN2-203) and two miRNAs (miR-325-3p and miR-384-5p) in this transition. Through the use of in vitro mESC differentiation and single-cell sequencing of CRISPR/Cas9 mutants, we demonstrate that lncMN2-203 affects MN differentiation by sponging miR-466i-5p and upregulating its targets, including several factors involved in neuronal differentiation and function. In parallel, miR-325-3p and miR-384-5p, co-transcribed with lncMN2-203, act by repressing proliferation-related factors. These findings indicate the functional relevance of the MN2 locus and exemplify additional layers of specificity regulation in MN differentiation.

中文翻译：

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一个多功能基因座通过短和长的非编码 RNA 控制运动神经元的分化

从分裂祖细胞到有丝分裂后运动神经元 (MN) 的转变是由一系列事件协调的，这些事件主要通过分析特定编程转录因子的活性在转录水平上进行研究。在这里，我们确定了在这种转变中含有 lncRNA (lncMN2-203) 和两个 miRNA (miR-325-3p 和 miR-384-5p) 的 MN 特异性转录单位 (MN2) 的转录后作用。通过使用体外mESC 分化和 CRISPR/Cas9 突变体的单细胞测序，我们证明 lncMN2-203 通过海绵化 miR-466i-5p 和上调其靶标（包括参与神经元分化和功能的几个因素）来影响 MN 分化。同时，与 lncMN2-203 共同转录的 miR-325-3p 和 miR-384-5p 通过抑制增殖相关因子起作用。这些发现表明了 MN2 基因座的功能相关性，并举例说明了 MN 分化中额外的特异性调节层。