What started out as a painstakingly slow project with trouble establishing a reliable experimental system has turned in to a very promising project. I have been able to show that expression of p16 influences the sensitivity of Schwann cell lines and MPNST cell lines to SIRT5 depletion. In the past week, I was able to corroborate these findings using Annexin V and propidium iodide staining followed by flow cytometry. I have also ordered lentivirus to perform knockdowns of p16 to see if this confers sensitivity to SIRT5 depletion.
Additionally I will be analyzing cell cycle stage specific phenotypes using the fluorescence ubiquitination cell cycle indicator system. This will also be done using flow cytometry so I will likely become very experienced at applying this method in the coming months.
Over the course of the Summer, I have become proficient at a number of new techniques. I learned primer design and CRISPR guide RNA design. I also gained a decent amount of experience in cloning. Recently, I learned the process of generating lentivirus starting from a bacterial glycerol stock. I also analyzed flow cytometry data for the first time. In the coming months I will likely submit the literature review I have been working on. I will also begin writing my honors thesis for CMB. I am looking forward to hearing back about a grant that I recently submitted and I’ll soon be applying for the NSF graduate research fellowship program. I’ve learned more this Summer than at any other point in my college career.