UNASSIGNED: We characterized circRNA-associated competitive endogenous RNA (ceRNA) networks in brain tissue of low folate -induced NTDs mouse at embryonic day 13.5 by high-throughput sequencing. The expression levels of Circzfp644, miR-20-5p and Gas7 were detected by RT-PCR. Gas7 and Circzfp644 functions were determined by miRNA-mimics and inhibitors in mouse teratocarcinoma cells (F9 cells), and luciferase gene reporter assay was assessed in the F9 cells. In addition, the expression levels of Circzfp644, miR-20-5p and Gas7 were determined by Nanostring in human NTDs tissues.
UNASSIGNED: We detected 57 circRNA transcripts, 16 miRNAs, and 148 mRNAs that were significantly dysregulated in NTDs brain tissues compared with their expression levels in control (normal) tissues. Circzfp644 shared miRNA response elements with the growth arrest specific 7 ( Gas7) gene and competitively bound with miR-20-5p to increase the expression of Gas7. Downregulation of Circzfp644 and Gas7 and upregulation of miR-20-5p were found in human NTD tissue.
UNASSIGNED: This study provides new perspectives on the role of circRNAs in nervous system development and the pathogenesis of NTDs.
■我们通过高通量测序表征了低叶酸诱导的NTDs小鼠在胚胎第13.5天的脑组织中与circRNA相关的竞争性内源性RNA(ceRNA)网络。RT-PCR检测Circzfp644、miR-20-5p和Gas7的表达水平。Gas7和Circzfp644的功能是通过小鼠畸胎瘤细胞(F9细胞)中的miRNA模拟物和抑制剂确定的,在F9细胞中评估荧光素酶基因报告基因测定。此外,通过Nanostring测定人NTDs组织中Circzfp644、miR-20-5p和Gas7的表达水平。
■我们检测到57个circRNA转录本,16个miRNA,与对照(正常)组织中的表达水平相比,NTDs脑组织中的148个mRNA明显失调。Circzfp644与生长停滞特异性7(Gas7)基因共享miRNA反应元件,并与miR-20-5p竞争性结合以增加Gas7的表达。在人NTD组织中发现了Circzfp644和Gas7的下调和miR-20-5p的上调。
■这项研究为circRNAs在神经系统发育和NTDs发病机理中的作用提供了新的视角。