Epi-CAP project, funded by Science Fund of the Republic of Serbia (Serbian Science and Diaspora Collaboration Program: Knowledge Exchange Vouchers), represents a continuation of previous research done within the Department of Molecular Biology, IBISS in which Epi-CRISPR constructs was used for targeted DNA methylation. By targeting defined promoter sequence of the homeobox Arx gene in alpha-cells, trans-differentiation of mouse pancreatic alpha- to insulin-producing cells (Epi-cells) was achieved. The next step in the research was to compare chromatin landscapes of control alpha- and beta-cells and Epi-cells by using ATAC-seq methodology in order to further enhance understanding of the epigenetic maintenance mechanisms that define the phenotypes of the mouse pancreatic alpha-, beta- and Epi-cell. In Serbia, alpha-cells were transfected with Epi-CRISPR construct (deactivated Cas9 fused to DNMT3a3L and 4 different guided RNA for targeting Arx gene) and sorted, after which Epi-cells including negative (alpha-cells) and positive (beta-cells) controls were shipped in triplicate to the host laboratory were ATAC-seq is routinely used. Upon arrival of the Serbian counterpart to USA, the protocol for ATAC-seq was optimized for animal cells to obtain the best quality and quantity of sequencing library.

The lecturers are:
Dr. Melita Vidaković, Principal Research Fellow at Department of Molecular Biology IBISS (PI)

Dr. Paja Šijačić, Postdoctoral Fellow at Department of Biology, Emory University, Atlanta, USA (Project Partner - expert from diaspora)

Dr. Anja Tolić, Research Associate at Department of Molecular Biology IBISS

Dr. Jovana Rajić, Research Associate at Department of Molecular Biology IBISS

This presentation is available at the IBISS YouTube channel.