High-content microscopy platform for multiplex perturbation/multiplex readout studies

University Of Tx Md Anderson Can Ctr, Houston TX

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Abstract

NOT-GM-24-021 – ABSTRACT – HART Highly multiplexed genetic perturbations offer huge potential beyond fitness screens. High throughput microscopy offers a tractable method for combining complex genetic perturbation with multiparameter functional assays. Here we propose the purchase of a Nikon Eclipse Ji microscopy system to facilitate complex data collection to decipher the complexity and redundancy of mammalian signaling pathways. Using our next- generation CRISPR/Cas12a polygenic knockout system, we combine custom CRISPR arrays targeting signaling effectors and cell surface “barcodes”, then use immunofluorescence readouts of cell state across dozens of targets – two to four per well in a 96 well plate -- to understand the dynamics of cell signaling.