Biomaterial Database

Single particle tracking assay to study coronavirus membrane fusion. 2015

Deirdre A Costello, and Susan Daniel

School of Chemical and Biomolecular Engineering, Cornell University, 120 Olin Hall, Ithaca, NY, 14853, USA.

Single particle tracking (SPT) of individual virion fusion with host cell membranes using total internal reflection microscopy (TIRFM) is a powerful technique for quantitatively characterizing virus-host interactions. One significant limitation of this assay to its wider use across many types of enveloped viruses, such as coronavirus, has been incorporating non-lipid receptors (proteins) into the supported lipid bilayers (SLBs) used to monitor membrane fusion. Here, we describe a method for incorporating a proteinaceous viral receptor, feline aminopeptidase N (fAPN), into SLBs using cell blebbing of mammalian cells expressing fAPN in the plasma membrane. This receptor binds feline coronavirus (FECV 1683). We describe how to carry out single particle tracking of FECV fusion in this SLB platform to obtain fusion kinetics.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D008051	Lipid Bilayers	Layers of lipid molecules which are two molecules thick. Bilayer systems are frequently studied as models of biological membranes.	Bilayers, Lipid,Bilayer, Lipid,Lipid Bilayer
D008561	Membrane Fusion	The adherence and merging of cell membranes, intracellular membranes, or artificial membranes to each other or to viruses, parasites, or interstitial particles through a variety of chemical and physical processes.	Fusion, Membrane,Fusions, Membrane,Membrane Fusions
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D006224	Cricetinae	A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.	Cricetus,Hamsters,Hamster
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D013194	Staining and Labeling	The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts.	Histological Labeling,Staining,Histological Labelings,Labeling and Staining,Labeling, Histological,Labelings, Histological,Stainings
D014771	Virion	The infective system of a virus, composed of the viral genome, a protein core, and a protein coat called a capsid, which may be naked or enclosed in a lipoprotein envelope called the peplos.	Virus Particle,Viral Particle,Viral Particles,Particle, Viral,Particle, Virus,Particles, Viral,Particles, Virus,Virions,Virus Particles
D016765	Coronavirus, Feline	A species of CORONAVIRUS infecting cats of all ages and commonly found in catteries and zoos. Cats are often found carrying the virus but only a small proportion develop disease. Feline coronavirus and Feline infectious peritonitis virus (FIPV) are virtually the same virus in genetic and antigenetic terms, and are morphologically indistinguishable. Since they only differ in their disease potential (with FIPV causing a more serious illness), they are considered biotypes of each other.	Coronavirus, Feline Enteric,FECV,FIPV,Feline Enteric Coronavirus,Feline infectious peritonitis virus,Infectious Peritonitis Virus, Feline,Peritonitis Virus, Infectious Feline,Coronaviruses, Feline,Coronaviruses, Feline Enteric,Enteric Coronavirus, Feline,Enteric Coronaviruses, Feline,Feline Coronavirus,Feline Coronaviruses,Feline Enteric Coronaviruses
D044085	Microfluidics	The study of fluid channels and chambers of tiny dimensions of tens to hundreds of micrometers and volumes of nanoliters or picoliters. This is of interest in biological MICROCIRCULATION and used in MICROCHEMISTRY and INVESTIGATIVE TECHNIQUES.	Microfluidic