Research Department: Internal Medicine-Cardiology Graduation Date: December 2019
Abstract: The recent developing attention toward translationally regulated gene expression has led many to pursue next-generation sequencing due to the low amount of obtainable starting material. Of the many modern sequencing technologies, ribosome profiling, RNA-sequencing, and ATAC-sequencing are commonly used sequencing technologies for the interpretation of gene expression via the analysis of mRNA, nRNA, and DNA, respectively. Ribosome profiling, in particular, has been used to determine the position of a ribosome and measure the translational rates of a cell, a prime factor in gene expression. Unlike polysome profiling and TRAP-sequencing, ribosome profiling only sequences the translatome, or the actively translated mRNA regions of the cell, from all cell types. The sequenced ribosome-protected mRNA fragments are able to be mapped back to coding sequences. In our lab, we are studying nuclear isolation from all cell sub-types in mouse heart tissue. This project is guided towards partitioning out the ribosomes believed to be located on the perinuclear membrane from our nuclei isolation protocol components via density centrifugation. The protocol was first tested on HEK293 kidney cells to verify the location of the ER and the ribosomes in the pelleted components. Once verified, the protocol will be optimized toward cycloheximide treatment of mammalian tissue. Overall, we seek to design a protocol to produce mid-translational mRNA for ribosome profiling along with nuclear RNA for RNA-sequencing and DNA for ATAC-sequencing from the same sample.
What does research mean to you? Research is a collaboration spanning years of dedicated work guided toward one common goal: the betterment of human life. The research I have seen and conducted have all been the result of a team effort, and I believe that we have progressed so far because research is not treated as a one-man sprint but rather as a relay race. The work of a single person can’t match the work of the many. Tell us about your journey. During my last two years of high school, I attended ASMSA (similar to TAMS), a boarding school/ junior college for science, math, and the arts. A major requirement for graduating from ASMSA was to complete a 1.5-year research project. For most of my life until 2 years ago, I guided my life toward practicing medicine, so the project seemed trivial to me. However, I wanted to give it my best. I was accepted into a summer research position at UAMS and placed on a project that studied the expression of the CRTAP/P3H1 Osteogenesis Imperfecta-associated complex in E. Coli DH5α. The experiences I gained from my time at UAMS conducting research changed my entire outlook on research as a whole. I no longer saw it as a mind-bending and lonely task but rather as a supported challenge that caused me to grow further in not only my education but also in my passion for the wellbeing of others. From then on, I knew that no matter what I did, research had to be a large component of my life. During my time as a Green Fellow in the lab of Dr. Nikhil V. Munshi, M.D., Ph.D., I faced many issues ranging from failed trials to failed hypotheses, but the advice I gained from my lab helped me achieve more than I thought possible. Research consumed most of my daily life, but I loved the process. I was reading multiple papers that encompassed research outside my project, giving me a better view of proper research technique, verification, and presentation. The Munshi lab was an inviting community that fostered learning and an openness to ideas. This experience shaped my freshman experience and molded me into a more well structured student and researcher, for which I am grateful. The best advice I received from my PI was that research is 99% failure and 1% success. That 1% is achieved through hard work and humility. You must be willing to accept and provide help and support.
Advice for Future Green Fellows
Research is not simple, so do not see a failure as a stopping point, but as a chance for redirection. Do not let yourself become distraught by a failure but grow from it. Grow from your failures and criticisms. Learn to love the process of research more than the results. Enjoy the failures as much as the success, and in the end, it all leads to solid research and development. If you would like further advice regarding the Green Fellowship, I would be more than happy to help.