Genome contacts of KLF5 gene promoter region in pancreatic ductal adenocarinoma cells

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and chemotherapy resistant tumors with high metastatic potential. The mechanisms of PDAC progression are not sufficiently studied. It is known that during tumor progression the spectrum of the expressed transcription factors is changed substantially. In particular, with the increase of the tumor grade, expression level of KLF5 gene decreases. The aim of the project is to search the genome for distant KLF5 regulatory elements which activity is related to the PDAC grade, and to reveal factors that control the activity of these genes. The search will be performed using the circular chromosome conformation capture with high throughput sequencing (4C-seq) using HinDIII and DpnII restriction endonucleases. We intend to reveal the chromatin regions which are distant from KLF5 promoter region in linear genome but are close to these sequence in the three-dimensional nuclear structure in cell lines which simulate the main stages of the PDAC progression. These regions are highly likely to contain tissue specific enhancers and/or silencers of corresponding gene and other regulatory regions as well. Next the properties of these regions, such as their chromatin structure, regulatory proteins binding, the activity of these sequences in the reporter gene assay and the comparison of position of these regions with known structural elements of the genome (e.g. topologically associated domains, contact domains etc.) should allow us to identify regulatory pathways participating in the progression of PDAC and involvement in this process of KLF5 transcription factor.