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Open AccessArticle

Fluorescent Protein Inserts in between NC and SP2 Are Tolerated for Assembly, Release and Maturation of HIV with Limited Infectivity

by Mourad Bendjennat 1,2,3,* and Saveez Saffarian 2,3,4,*
1
Radiation Oncology Department, University of Miami Miller SOM-SCCC, Miami, FL 33136, USA
2
Department of Physics and Astronomy, University of Utah, Salt Lake City, UT 84112, USA
3
Center for Cell and Genome Science, University of Utah, Salt Lake City, UT 84112, USA
4
Department of Biology, University of Utah, Salt Lake City, UT 84112, USA
*
Authors to whom correspondence should be addressed.
Viruses 2019, 11(11), 973; https://doi.org/10.3390/v11110973
Received: 5 September 2019 / Revised: 8 October 2019 / Accepted: 14 October 2019 / Published: 23 October 2019
We report the design of a fluorescent HIV construct that is labeled by insertion of fluorescent protein between the nucleocapsid (NC) and spacer peptide 2 (SP2) domains of Gag and further show that the fluorescent protein is released from its confines within Gag during maturation. This fluorescent HIV is capable of budding and maturation with similar efficiency to the parental virus. Virions generated using this design within the R8 HIV backbone pseudotyped with VSV-G were capable of delivering small RNA genomes encoding GFP to the target cells; however, the same design within the NL4-3 backbone has limited HIV infectivity. The virions generated by these constructs are approximately 165 ± 35 nm in size, which is significantly larger than wild type HIV. We suggest that this design has the potential to be a vehicle for protein and small guide RNA delivery. View Full-Text
Keywords: fluorescent HIV; NL4-3(dGFP); R8(dGFP); NL4-3(iGFP) fluorescent HIV; NL4-3(dGFP); R8(dGFP); NL4-3(iGFP)
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Bendjennat, M.; Saffarian, S. Fluorescent Protein Inserts in between NC and SP2 Are Tolerated for Assembly, Release and Maturation of HIV with Limited Infectivity. Viruses 2019, 11, 973.

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