Tracking and quantitation of fluorescent HIV during cell-to-cell transmission.
The green fluorescent protein (GFP) is a powerful genetic marking tool that has enabled virologists to monitor and track viral proteins during HIV infection. Expression-optimized Gag-GFP constructs have been used to study virus-like particle (VLP) assembly and localization in cell types that are easily transfected. The development of HIV-1 variants carrying GFP within the context of the viral genome has facilitated the study of infection and has been particularly useful in monitoring the transfer of virus between cells following virological synapse formation. HIV Gag-iGFP, a viral clone that contains GFP inserted between the matrix (MA) and capsid (CA) domains of Gag, is the first replication competent molecular clone that generates fluorescent infectious particles. Here, we discuss some methods that exploit HIV Gag-iGFP to quantify cell-to-cell transmission of virus by flow cytometry and to track the proteins during assembly and transmission using live-cell imaging.
Dale
Benjamin M
BM
Division of Infectious Diseases, Department of Medicine, Immunology Institute, Mount Sinai School of Medicine, New York, NY 10029, USA.
McNerney
Gregory P
GP
Hübner
Wolfgang
W
Huser
Thomas R
TR
Chen
Benjamin K
BK
D058948:Cell Tracking / Q000379:methods*; D003588:Cytopathogenic Effect, Viral; D005434:Flow Cytometry; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000235:genetics*; D054884:Host-Pathogen Interactions; D006801:Humans; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D030781:Organisms, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis / Q000235:genetics
D011993:Recombinant Fusion Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins