Abstract by Alec Wallace
Chemistry and Biochemistry
Developing a tool for CMG2 proximity proteomics: Cloning a CMG2-EGFP-BirA knock-in using CRISPR
Angiogenesis, the process of forming new blood vessels, plays a significant role in various pathologies including tumor growth and metastasis. Capillary Morphogenesis Gene 2 (CMG2) has been identified as an important regulator of angiogenesis and as a target for new therapeutics. However, little is known about the intracellular pathways of CMG2. Our lab has performed preliminary proximity-dependent biotinylation (BioID) experiments by overexpressing a CMG2-BirA construct and identified potential interactors. To make these experiments more physiologically relevant, we are developing a CMG2-EGFP-BirA knock-in using Gibson assembly and CRISPR technology to allow the proximity proteomics experiment to be performed under the control of the native CMG2 promotor and in the biologically relevant endothelial cell background. Here we report our progress in developing this tool.