Optimizing Viral Detection: Creating Fluorescent Chikungunya Virus Infectious Clones

Technical Report · Fri Jul 01 04:00:00 EDT 2016

DOI:https://doi.org/10.2172/1561803· OSTI ID:1561803

Kaszubinski, Sierra ^[1]

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1. Sandia National Laboratories (SNL-CA), Livermore, CA (United States)

Human infection with Chikungunya virus (CHIKV) results in debilitating joint pain and arthritis that can persist for months or even years. CHIKV is a re-emerging virus responsible for large epidemics in Asia, Africa, Europe, and most recently South and Central America. Yet, CHIKV has no specific treatment or vaccine. Therefore, steps towards eradicating the virus are important to public safety. Tools to facilitate drug screens are especially important when searching for viable treatments. Therefore, our lab sought to create reporter CHIKV encoding fluorescent markers, to detect infection in cells. Cloning was used to develop a CHIKV infectious clone encoding GFP. Creation of this tool required use of PCR, Gibson assembly, and the CRISPR Cas9 system. We are also working to create a CHIKV expressing mCherry. Next steps include determining the efficiency of these infectious clones compared to wildtype virus and performing drug screen studies. I also made contributions to other projects including development of a CRISPR Cas9 guide RNA library, a bioinformatics project using new software, and CRISPR Cas9 expressing cell line, that will be used in Zika virus studies. Finally, I assisted in development of a market survey focused on biodefense technologies for DHS.

• Research Organization: Sandia National Laboratories (SNL-CA), Livermore, CA (United States)
• Sponsoring Organization: USDOE National Nuclear Security Administration (NNSA)
• DOE Contract Number: AC04-94AL85000
• OSTI ID: 1561803
• Report Number(s): SAND--2016-7246R; 646185
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES