Identification of Grhl3 Target Genes During the Mesenchymal to Epithelial Transition
Ahmad Saleh Zeid, Diana
An Najah National University
The epithelial to mesenchymal transition (EMT) and the mesenchymal to epithelial transition (MET) are essential processes that occur throughout embryonic development and are critical for organogenesis and morphogenesis, resulting in dynamic changes in cell identity. The transcription factors Grhl3 and other regulatory genes play a key role in regulating E-cad expression during the MET process, which is a hallmark of the procedure. This work aims to identify Grhl3's transcriptional targets during MET and assess their potential relevance. To test the hypothesis that a transcriptional network that controls the stability of the epithelial state controls the dynamic regulation of MET and that this network may also play a role in the initiation of MET, Analysis of RNA sequencing data containing 14055 genes differentially expressed genes was carried out using pipelines for RNA sequencing analysis such as DESeq2 to identify Grhl3 target genes during MET bioinformatically. Then, using a loss of function approach, where Grhl3 was downregulated by siRNAs, putative Grhl3 target genes were validated by qPCR. The results showed that cells could not initiate MET when Grhl3 was silenced. There is a correlation between the expression of the knocked-down Grhl3 and the chosen target genes in both RNA-Seq results and the experimental results, where they all were downregulated. Cenpf was the most downregulated with the Grhl3 RNA interference from the RNA-Seq. On the other hand, Knl1 was the most downregulated experimentally. These findings suggest that Grhl3 is one of the essential transcription factors that direct the MET process, as well as that Grhl3, has many target genes that are differentially correlated with its gene expression and that Hmgb2, Nusap1, Cenpf, and Knl1 could be good candidates as Grhl3 target genes, which should be further investigated and more experiments carried out to emphasize the results.
mesenchymal , morphogenesis , organogenesis