With Keck Foundation support, Colorado State researchers to study how viruses attack

August 30, 2016

FORT COLLINS, COLORADO - Within the tiniest of spaces - single molecules in single cells - Colorado State University scientists are aiming for something huge: illuminating in never-before-seen detail exactly how viruses hijack their host cells.

Life science researchers Tim Stasevich and Brian Musky have received a four-year, $1.2 million grant from the W.M. Keck Foundation for a project that combines sensitive microscopes and sophisticated computation to quantify protein expression in single cells. Their project is titled "Multiplexed Real-Time Quantification of RNA to Protein Translation in Live Cells," and is bolstered by additional support from CSU.

Stasevich is an assistant professor in the College of Natural Sciences' Department of Biochemistry and Molecular Biology, and Munsky is an assistant professor in the College of Engineering's Department of Chemical and Biological Engineering and in the School of Biomedical Engineering. Together, they are seeking to quantify the translation and expression of up to 100 different RNA transcripts in real time, in living cells.

Stasevich and Munsky both started at CSU as assistant professors in 2014, and quickly discovered the intersection of their research goals. Stasevich, an experimentalist, develops microscopy and protein tagging techniques to image real-time cellular processes, including most recently, RNA translation. Munsky is a computational theorist who applies statistical tools to discriminate and characterize extremely subtle and seemingly random processes within single cells.

"We are grateful to the Keck Foundation for supporting our work, which we hope will have a profound impact on the fundamental understanding of gene expression networks," Stasevich said. "Our study will offer new insight into some longstanding mysteries surrounding RNA translation and viral biogenesis."

In the Keck project, Stasevich's imaging of real-time RNA translation at the single-molecule level will be amplified by computationally "fingerprinting" brightness fluctuations of tagged proteins as they're translated. By understanding the dynamics of these rapid-fire signals, Munsky can help differentiate and characterize many distinct signals, and this analysis could lead to a better understanding of all the dynamics of gene expression.

"We can make use of statistical fluctuation information to go beyond the limited color pallet of protein tag technologies," Munsky said. "We hope to be able to identify dozens or hundreds of different biological signals."

For the Keck proposal, Stasevich and Munsky will study a phenomenon called ribosomal frame shifting, a process that viruses use to encode more than one protein on the same region of DNA or RNA. Viruses' genomes are thus very small, allowing them to efficiently take over a host cell by hijacking the cell's RNA translation machinery.

Stastevich's microscopy can illuminate this ribosomal frame shifting in real time. Supported by computational analysis of each step in this shifting process, the researchers seek to predict and control how viruses hijack translational machinery in cells in order to replicate themselves. This is a process that's been studied, but never before seen.

The Keck Foundation grant also provides for the establishment of the W.M. Keck Gene Imaging Lab and the W.M. Keck High Performance Computing Cluster. Both of these facilities will be of central importance to the success of Stasevich and Munsky's project.

The Keck Foundation has provided seven grants to CSU scientists in the last 15 years. Most recently, chemists Amy Prieto and Jamie Neilson were supported for their project on novel materials discovery.

The Los Angeles-based Keck Foundation was established in 1954 by the late W. M. Keck, founder of the Superior Oil Company. The foundation's grant-making is focused primarily on pioneering efforts in the areas of medical research, science and engineering and undergraduate education. The foundation also maintains a Southern California Grant Program that provides support for the Los Angeles community, with a special emphasis on children and youth. For more information, visit http://wwwwmkeck.org.
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Colorado State University

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