Description: CRISPR (clustered regularly interspaced short palindromic repeats), is a cutting-edge technology, which is revolutionizing how to treat and diagnose diseases. The recent FDA approval of a CRISPR therapeutic for the treatment of sickle cell anemia indicates that we will be seeing more CRISPR therapeutics on the horizon. Adhesion proteins play a crucial role in cell-cell adhesion, cell-substrate adhesion, immune responses, signal transduction, and wounding healing. Students will design sgRNA molecules to target adhesion genes in a canine macrophage-like cells and generate knockouts. For this CURE lab, we will be pairing with the animal health biotechnology company, Zoetis. Zoetis employees will be sharing their expertise on CRISPR, cell culture, and flow cytometry.
Skills Gained: Basic cell culture skills such as thawing frozen cell stocks, passaging, feeding cells, and making complete cell culture media. Additional skills include PCR, running agarose cells, sgRNA guide design, transfection, single cell clonal dilution, extraction and quantification of DNA, interpretation of Sanger sequencing results to confirm CRISPR edits, and flow cytometry to confirm edit of cell surface markers.