|Date: Monday, November 13, 2017
Location: 335 West Hall (12:00 PM to 1:00 PM)
Title: Real-time imaging of single mRNA translation dynamics in living cells
Abstract: We are developing technology to image single RNA translation dynamics in living cells. Using high-affinity antibody-based probes, multimerized epitope tags, and single molecule microscopy, we are able to visualize and quantify the emergence of nascent protein chains from single pre-marked RNA1. Here, I'll describe this technology as well as a two color extension useful for comparing translation rates between two different parts of a single open reading frame (ORF) or two different ORFs. Using information from the correlations of fluorescence fluctuations, we can accurately quantify single mRNA translation elongation rates in both tagged and untagged portions of ORFs. By transiently loading probes and reporter DNA into cells in a combinatorial fashion, multiplexed imaging of gene expression is possible. Preliminary application of this technology to the study of viral frameshifting will be discussed.
Speaker: Tim Stasevich
Institution: Colorado State University
Event Organizer: Victoria Booth firstname.lastname@example.org