Using the RNA Arrayed-Hybrid Purification (RAP) Method to Study the Novel lincRNA Firre

Mentors: Jesse Engreitz, Mitch Guttman

Ben’s research focused on large intergenic non-coding RNAs (lincRNAs), which play important roles in diverse cellular processes, but do not encode proteins. The goal of Ben’s project was to examine the function of the lincRNA Firre. Firre has been shown to localize to both X chromosomes, and is necessary for adipocyte (fat cell) differentiation. Understanding adipogenesis is important for elucidating the mechanism behind metabolic diseases such as diabetes.

Ben utilized a new method called RNA Arrayed-Hybrid Purification (RAP) to determine where in the genome the lincRNA Firre binds. To perform the RAP method, Ben used cross-linked cells, to preserve RNA-DNA-protein complexes that exist in those cells. Next, he synthesized complementary RNA probes to Firre, which he then used to pull-down and isolate complexes containing Firre from cell lysates. Ben and his mentors then sequenced the DNA that was complexed with Firre, in order to identify the DNA sites to which Firre binds.



Ben, a senior at Brookline High School, used a new method to study non-coding RNAs in cells, focusing on a lincRNA called Firre, which is necessary for fat cell differentiation.