Generation of hiPSC reporter lines to study the development of endocrine cells
Alpha, beta and delta cells are endocrine cells of the pancreas and play a fundamental role in the regulation of blood glucose levels. However, how the individual cell types develop from a common precursor is still relatively poorly understood. In order to better understand the specification of individual endocrine cell types, we are working in this project on the generation of gene-modified cell lines that can be used to observe individual developmental steps in real time. To this end, double-strand breaks are first induced in relevant genes in human induced pluripotent stem cells (hiPSCs) using CRISPR/Cas9. The coding sequence of the reporter gene, for example a fluorescent protein, is then inserted. This is done during the DNA repair process by homology-directed repair (HDR) and the provision of an artificial repair template. The cell lines generated in this way ("reporter lines") should in future provide information about the temporal sequence of individual developmental processes of endocrine cells and show ways in which these can be manipulated.
Project duration: 2023 - 2024
