MiR-124 synergism with ELAVL3 enhances target gene expression to promote neuronal maturity [HNs_RNAseq]

Neuron-enriched microRNAs (miRNAs), miR-9/9* and miR-124 (miR-9/9*-124), direct cell fate switching of human fibroblasts to neurons when ectopically expressed by repressing anti-neurogenic genes. How these miRNAs function after the onset of the transcriptome switch to a neuronal fate remains unclear. Here, we identified direct targets of miRNAs by Argonaute (AGO) HITS-CLIP as reprogramming cells activate the neuronal program and reveal the role of miR-124 that directly promotes the expression of its target genes associated with neuronal development and function. Overall design: MiR-124 and ELAVL3 activities were knockdown using a tough decoy and shRNA, respectively, in primary human neurons. Duplicates for control (TuD-miR-NS + shCTL) and knockdown (TuD-miR-124 + shELAVL3) were collected for RNAseq.

神经元富集的微RNA（microRNAs，miRNAs）miR-9/9*与miR-124（合称miR-9/9*-124）在异位表达时，可通过抑制抗神经生成基因，直接将人成纤维细胞的细胞命运转换为神经元细胞。目前，这类微RNA在转录组启动向神经元命运转换后如何发挥功能仍不明确。本研究在重编程细胞激活神经元程序的过程中，通过Argonaute（AGO）HITS-CLIP技术鉴定了miRNAs的直接靶标，并揭示了miR-124可直接促进其与神经元发育及功能相关靶基因的表达。实验设计概述：本研究分别采用刚性诱饵（tough decoy，TuD）与短发夹RNA（shRNA）敲低原代人神经元中miR-124与ELAVL3的活性；设置对照组（TuD-miR-NS + shCTL）与敲低组（TuD-miR-124 + shELAVL3）各设置生物学重复，收集样本进行RNA测序（RNAseq）。