College of Veterinary Medicine, Jilin Agricultural University, Changchun, China.
Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, Changchun, China.
Front Cell Infect Microbiol. 2021 Dec 9;11:806290. doi: 10.3389/fcimb.2021.806290. eCollection 2021.
In previous experiments, we identified the effect of deletion of the Zbtb1 gene on circRNAs and microRNAs. In this study, we examined the expression profiles of lncRNAs and mRNAs using the RNA-seq method for Zbtb1-deficient EL4 cells and performed a clustering analysis of differentially expressed lncRNAs and mRNAs. GO term histograms and KEGG scatter plots were drawn. For the experimental results, a joint analysis was performed, which predicted the regulatory relationships among lncRNAs, mRNAs, microRNAs and circRNAs. For the regulatory relationship between lncRNAs and target genes, the chromatin structure and the degree of openness were verified for the possible target gene locations regulated by lncRNA using experimental methods such as Hi-C and ATAC-seq. Ultimately, the possible differential regulation of the Brcal and Dennd5d genes by lncRNAs and the differential changes in transcription factor binding sites in the promoter region were identified. For neRNA-regulated target genes with significantly differentially expressed mRNAs, a combined screen was performed, and the final obtained candidate target genes were subjected to GO and KEGG term enrichment analyses. Our results illustrate that the Zbtb1 gene can not only function as a regulatory factor but also regulate EL4 cells from multiple perspectives based on ceRNA theory.
在之前的实验中,我们确定了删除 Zbtb1 基因对 circRNAs 和 microRNAs 的影响。在这项研究中,我们使用 RNA-seq 方法检查了 Zbtb1 缺陷型 EL4 细胞中的 lncRNAs 和 mRNAs 的表达谱,并对差异表达的 lncRNAs 和 mRNAs 进行聚类分析。绘制了 GO 术语直方图和 KEGG 散点图。对于实验结果,进行了联合分析,预测了 lncRNAs、mRNAs、microRNAs 和 circRNAs 之间的调控关系。对于 lncRNA 与靶基因的调控关系,使用 Hi-C 和 ATAC-seq 等实验方法,验证了 lncRNA 可能调控的靶基因位置的染色质结构和开放程度。最终,确定了 lncRNA 对 Brcal 和 Dennd5d 基因的可能差异调控以及启动子区域转录因子结合位点的差异变化。对于具有显著差异表达 mRNAs 的 neRNA 调控靶基因,进行了联合筛选,最终获得的候选靶基因进行了 GO 和 KEGG 术语富集分析。我们的结果表明,Zbtb1 基因不仅可以作为调控因子发挥作用,而且还可以根据 ceRNA 理论从多个角度调节 EL4 细胞。