Intrabodies are engineered antibodies that work inside living cells, enabling a wide range of therapeutic, diagnostic, and imaging applications. In my lab, we use intrabodies to visualize the dynamics of protein post-translational modifications and mRNA translation—two processes that are notoriously difficult to capture by other means. In the first part of the talk, I’ll describe a new AI-driven pipeline that integrates AlphaFold2, ProteinMPNN, and live-cell screening to dramatically accelerate intrabody design. This approach has yielded dozens of functional scFv intrabodies targeting synthetic, viral, and histone epitopes for live imaging. In the second part, I’ll showcase how we’re applying these tools to track single mRNA translation events. Using new All Probes Plasmids (APPs) and photostable mStayGold reporters, we can image translation on individual mRNAs for hours and repeatedly measure elongation and initiation on the same transcript. Together, our designer intrabodies and optimized imaging tools lower the barrier to visualizing full protein lifetimes inside living cells.
https://stasevichlab.colostate.edu/
