Virus evolution and host responses in positive-sense RNA viruses

National Institute Of Allergy And Infectious Diseases

Investigators

Linked publications & trials

Abstract

In the first year of the QVEU, we have established three of the long-term projects in the lab. First, in work led by Visiting Fellow Dr. Nathania Dabilla, we are establishing single-cell sequencing approaches to observe the heterogeneous and dynamic process of infection in the complex selective environment in vivo. In the last year, we have developed the molecular techniques that will allow us to sequence consensus viral genomes in parallel with sequencing the host transcriptome in individual cells. Secondly, in work led by Visiting Fellow Dr. William Bakhache, we have developed our first mutational scanning libraries. Our approach, based on the Deep Insertional and Mutational Protein Library Engineering (DIMPLE) technology, allows us to engineer complete libraries of insertions, deletions, and coding substitutions into every position of a protein. We are currently characterizing our first of these libraries focused on the capsid region of the human enterovirus, EV-A71. Finally, in the past year we have established a computational and experimental pipeline to reconstruct the evolutionary histories and ancestral sequences of viral proteins. Our current studies are focused on the reconstruction of the historical evolution of structural proteins of flaviviruses, including the ancestors of dengue, Zika, and yellow fever virus. We have reconstructed and synthesized the ancestral sequences and are currently developing in biochemical and cell biological assays to characterize the function of these reconstructed ancestral proteins.