LRRK2: What are you up to?

On my very first day my amazing PI, Dr. Sanders, handed me a sheet of paper with three goals for this summer. I was very surprised that I was able to start working on my first goal, learning mammalian cell culture and proper aseptic technique that same day. Today, I am incredibly happy to say that I have completed that first goal. I have worked with a line of human embryonic kidney cells (HEK293). I can now successfully and independently (much to my mentor’s delight) split, count, thaw, make media for, and collect protein from these cells.

I will be using my mammalian cell culture techniques throughout the rest of the summer in an effort to meet my two other goals and to answer important scientific questions. One of my goals is to determine if HEK293 cells lacking a  LRRK2 gene are more sensitive to environmental toxicants and DNA damaging agents. To do this, I am planning to treat wild type cells with LRRK2 and cells that have had the LRRK2 gene removed with the same doses of certain toxicants and DNA damage inducing agents and analyze the differences. Some of the many common toxicants I am planning to use are rotenone, hydrogen peroxide, and mitomycin C. If I am able to complete this project and get promising results (fingers crossed), I can move on to my third and final goal.

My last goal is to determine whether LRRK2 deficiency leads to cell cycle checkpoints in the presence or absence of the same environmental and DNA damaging agents that were used to determine cell viability before. I will be checking the levels of unique proteins associated with different parts of the cell cycle to see if the checkpoints are working properly. I will be using DC protein assays and Western Blots in order to carry out these goals, as well as my mammalian cell culture technique to keep my cells happy throughout.

Why is this my project? My lab, the Sanders lab, has discovered the importance of mitochondrial DNA damage in neurons affected in Parkinson’s disease. The LRRK2 gene is known to be a genetically significant factor for the onset of Parkinson’s disease. Although scientists know that it plays a role in many Parkinson’s patients, they do not fully understand what it does. My lab believes that LRRK2 may play a crucial role in the mitochondrial DNA damage they found earlier and may play a critical role in the cell cycle when such DNA damage (produced by the exposures in vitro) is present. This is why I am looking at both cell viability and cell cycle checkpoints in HEK 293 cells with the LRRK2 gene and the same cells without the LRRK2 gene. We hope to understand this gene better, so we can understand how it is affecting Parkinson’s patients and to try and find a way to reverse the damage it does.

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