Generation of Functional Endocrine Cell Reporter Pluripotent Cell Lines
Contact PI: Paul Gadue, PhD
Recent advances in the in vitro differentiation of human pluripotent and iPS cells now allow the generation of mono-hormonal beta-like cells that can mature into functional beta cells in rodent models within a few weeks after transplantation with limited in vitro function. These protocols still generate a subpopulation of non-functional poly-hormonal population difficult as most functional assays examine the bulk response given stimuli. Therefore, we propose to generate reporter iPS cell lines that express reporter constructs in the INS, GCG, and SST loci using CRISPR/Cas9 genome editing technology. We propose to target a green fluorescent protein calcium indicator and a luciferase/pro-insulin fusion protein into the SST locus. The use of the luciferase/pro-insulin fusion protein offers advances n cost, ease of use, and potential sensitivity compared to c-peptide ELISAs that are currently used to assay insulin secretion.
Opportunity Pool Project Sponsored by CHIB.