Sean Hilgendorf, COL ’24, Howard Beach, NY
This was the second summer I had the opportunity to conduct research at Penn in the Brian Gregory Lab. I had the pleasure of working alongside my mentor, Wil Prall, to process and begin a phenotyping experiment on the Arabidopsis thaliana plant. The Arabidopsis plant is the model organism for plant biology, as it is both easy to maintain and its whole genome is widely available for all members of the scientific community. With that being said, our research specifically focuses on N6-Methyladenosine, more commonly referred to as m6A, which is an important post-transcriptional regulator in eukaryotes. Many studies test the deletion and overexpression of this gene and its impact on translation and the phenotypes of species it is modified in. The current Arabidopsis model with an m6A overexpressor is the MTA line. Despite being the current and best model, many studies have proved it to be a very poor model.
This is where our research has come in. My mentor, Wil, believes he has found three genes that work together in the arabidopsis genome that regulate m6A functionality. We spent a majority of the last year and a half preparing multiple Arabidopsis plant lines that either overexpress or have portions of these genes removed. We also took time to cross plants so they have two or all three of the genes removed or overexpressed. Much of this preparation involved DNA transformation, in which exogenous DNA in the form of plasmids were introduced into bacteria cells. Once we managed to make the specific DNA we wanted to insert into the plants, we infected the premature flower buds with our bacteria in order for their seeds to contain the mutant DNA. After this stage we selected and planted seeds that glowed under certain wavelengths of light (due to a specific gene added alongside our mutant DNA), which served as an indicator that the seeds obtained the bacterial DNA. After the plants grew, we isolated DNA from their leaves in order to genotype and confirm transformation was successful once again. Unfortunately, we had to plant the seeds for multiple generations to obtain homogeneity.
This brings me to our research’s current stage. At the end of the summer, we planted 22 different Arabidopsis plant lines, which included our specific experimental lines along with controls such as the wild type, and current MTA model. As these plants grow, my mentor and I will observe their physical characteristics (leaf size, leaf shape, leaf hairs, development time, etc.) and hopefully find a trend in the data. Once we notice a trend, we will try to determine the cellular cause of this derivation from the wild-type and MTA model and uncover its significance. Although we have a lot of data left to obtain, by having the opportunity to work in the lab this summer due to the Career Service Summer Funding Grant, I have made considerable progress with my mentor in this project and have become closer to publishing my first official paper.
This is part of a series of posts by recipients of the 2022 Career Services Summer Funding Grant. We’ve asked funding recipients to reflect on their summer experiences and talk about the industries in which they spent their summer. You can read the entire series here.