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Abstract
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.
Highlights
Fundamental research into mechanism of HIV virus replication has uncovered a complex release and maturation machinery that leads to delivery of RNA genomes to host cells [1]
R8.2 is derived from the full length HIV-1 R9 vector and incorporates all components of R9 except
Fundamental research in packaging and genome delivery of HIV had over the past 20 years led to development of potent life technology tools including the lentiviral delivery systems that are commonly used
Summary
Fundamental research into mechanism of HIV virus replication has uncovered a complex release and maturation machinery that leads to delivery of RNA genomes to host cells [1]. This process has been successfully utilized in the design of lentiviral vectors [2]. Gag and Gag–Pol proteins form the immature HIV lattice on the inner leaflet of the viral membrane [3]. The immature viral lattice is subjected to specific processing by HIV protease to produce the mature infectious virion [9,10].
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