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Cell–Cell Fusion Assays to Study Henipavirus Entry and Evaluate Therapeutics

Part of the Methods in Molecular Biology book series (MIMB,volume 2682)


Henipaviruses include the deadly zoonotic Nipah (NiV) and Hendra (HeV) paramyxoviruses, which have caused recurring outbreaks in human populations. A hallmark of henipavirus infection is the induction of cell–cell fusion (syncytia), caused by the expression of the attachment (G) and fusion (F) glycoproteins on the surface of infected cells. The interactions of G and F with each other and with receptors on cellular plasma membranes drive both viral entry and syncytia formation and are thus of great interest. While F shares structural and functional homologies with class I fusion proteins of other viruses such as influenza and human immunodeficiency viruses, the intricate interactions between the G and F glycoproteins allow for unique approaches to studying the class I membrane fusion process. This allows us to study cell–cell fusion and viral entry kinetics for BSL-4 pathogens such as NiV and HeV under BSL-2 conditions using recombinant DNA techniques. Here, we present approaches to studying henipavirus-induced membrane fusion for currently identified and emerging henipaviruses, including more traditional syncytia counting-based cell–cell fusion assay and a new heterologous fluorescent dye exchange cell–cell fusion assay.

Key words

  • Membrane fusion
  • Henipavirus
  • Class I fusion protein
  • Heptad repeat
  • Entry kinetics
  • Luciferase reporter assay
  • Split GFP
  • Split protein
  • F-triggering
  • Syncytia

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Correspondence to Hector C. Aguilar .

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Monreal, I.A., Aguilar, H.C. (2023). Cell–Cell Fusion Assays to Study Henipavirus Entry and Evaluate Therapeutics. In: Freiberg, A.N., Rockx, B. (eds) Nipah Virus. Methods in Molecular Biology, vol 2682. Humana, New York, NY.

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  • Print ISBN: 978-1-0716-3282-6

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