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为筛选在鸡胚胎干细胞(c ESCs)和鸡胚成纤维细胞(CEFs)中差异表达的micro RNA(miRNA),并探讨miRNA在维持c ESCs多能性过程中的作用,分别体外培养c ESCs、CEFs,应用miRNA芯片检测c ESCs、CEFs的miRNA表达谱,筛选差异表达的miRNA,并采用实时荧光定量PCR进行验证,同时利用生物信息学软件对靶基因进行预测。结果显示,从已知的993条鸡miRNA中,筛选出gga-miR-1777、gga-miR-302d、gga-miR-1607、gga-miR-1599和gga-miR-1576等19条在c ESCs中高水平表达的miRNA。实时荧光定量PCR检测结果显示,gga-miR-1777、gga-miR-302d的表达与miRNA芯片结果相符。生物信息学分析显示,gga-miR-1777、gga-miR-302d的靶基因中包含细胞分化相关的基因。研究结果提示,c ESCs中高水平表达的miRNA可能通过抑制分化基因的表达来维持c ESCs的多能性。
To screen differentially expressed microRNAs (miRNAs) in chicken embryonic stem cells (c ESCs) and chicken embryo fibroblasts (CEFs) and to explore the role of miRNAs in maintaining the pluripotency of c ESCs, culture c ESCs in vitro, CEFs were used to detect the miRNA expression profiles of c ESCs and CEFs using miRNA microarray. The differentially expressed miRNAs were screened and verified by real-time fluorescence quantitative PCR. Bioinformatics software was also used to predict the target genes. The results showed that 19 of 998 chicken miRNAs were screened for expression of g-miR-1777, gga-miR-302d, gga-miR-1607, gga-miR-1599 and gga-miR- High-level expression of miRNAs. Real-time quantitative PCR results showed that the expression of gga-miR-1777 and gga-miR-302d was consistent with the results of miRNA microarray. Bioinformatics analysis showed that the target genes of gga-miR-1777 and gga-miR-302d contained genes related to cell differentiation. The results suggest that high-level expression of miRNAs in c-ESCs may maintain the pluripotency of c-ESCs by inhibiting the expression of differentiated genes.