Lentiviral Vectors Pseudotyped with Filoviral Glycoproteins

Patrick L Sinn; Jeremy E Coffin; Natarajan Ayithan; Kathleen H Holt; Wendy Maury

doi:10.1007/978-1-4939-7116-9_5

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Journal article

Lentiviral Vectors Pseudotyped with Filoviral Glycoproteins

Patrick L Sinn, Jeremy E Coffin, Natarajan Ayithan, Kathleen H Holt and Wendy Maury

Methods in molecular biology (Clifton, N.J.), Vol.1628, pp.65-78

2017

DOI: 10.1007/978-1-4939-7116-9_5

PMCID: PMC5761728

PMID: 28573611

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https://www.ncbi.nlm.nih.gov/pmc/articles/5761728View

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Abstract

Pseudotyping lentivirus-based vectors is a strategy used to study conferred vector tropism and mechanisms of envelope glycoprotein function. Lentiviruses and filoviruses both assemble at the plasma membrane and have homotrimeric structural envelope glycoproteins that mediate both receptor binding and fusion. Such similarities help foster efficient pseudotyping. Importantly, filovirus glycoprotein pseudotyping of lentiviral vectors allows investigators to study virus entry at substantially less restrictive levels of biosafety containment than that required for wild-type filovirus work (biosafety level-2 vs. biosafety level-4, respectively). Standard lentiviral vector production involves transient transfection of viral component expression plasmids into producer cells, supernatant collection, and centrifuge concentration. Because the envelope glycoprotein expression plasmid is provided in trans, wild type or variant filoviral glycoproteins from marburgvirus or ebolavirus species may be used for pseudotyping and compared side-by-side. In this chapter we discuss the manufacture of pseudotyped lentiviral vector with an emphasis on small-scale laboratory grade production.

Genetic Therapy

Viral Envelope Proteins - genetics

Animals

Transfection

Humans

Plasmids - genetics

Lentivirus - genetics

Genetic Vectors

Virus Internalization

Membrane Glycoproteins - genetics

Viral Tropism - genetics

Details

Title: Subtitle: Lentiviral Vectors Pseudotyped with Filoviral Glycoproteins
Creators: Patrick L Sinn - Viral Vector Core Facility, University of Iowa, Iowa City, IA, USA
Jeremy E Coffin - Viral Vector Core Facility, University of Iowa, Iowa City, IA, USA
Natarajan Ayithan - Department of Microbiology, University of Iowa, Iowa City, IA, USA
Kathleen H Holt - Viral Vector Core Facility, University of Iowa, Iowa City, IA, USA
Wendy Maury - Department of Microbiology, University of Iowa, Iowa City, IA, USA. wendy-maury@uiowa.edu
Resource Type: Journal article
Publication Details: Methods in molecular biology (Clifton, N.J.), Vol.1628, pp.65-78
DOI: 10.1007/978-1-4939-7116-9_5
PMID: 28573611
PMCID: PMC5761728
NLM abbreviation: Methods Mol Biol
eISBN: 9781493971169; 1493971166
ISSN: 1064-3745
eISSN: 1940-6029
Grant note: R01 HL105821 / NHLBI NIH HHS P01 HL051670 / NHLBI NIH HHS P30 DK054759 / NIDDK NIH HHS P30 CA086862 / NCI NIH HHS R21 AI123616 / NIAID NIH HHS
Language: English
Date published: 2017
Academic Unit: Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics
Record Identifier: 9984083280602771

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