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Ebolaviruses pp 65–78Cite as

Lentiviral Vectors Pseudotyped with Filoviral Glycoproteins

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1628))

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.

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Acknowledgments

This work was supported by the National Institutes of Health R01 HL105821 (PLS) and R21 AI123616 (WM). The University of Iowa Viral Vector Core is partially supported by the Cystic Fibrosis Foundation and the National Institutes of Health: P30 DK054759, P01 HL051670, and P30 CA086862.

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Authors and Affiliations

  1. Department of Pediatrics, University of Iowa, Iowa City, IA, USA

    Patrick L. Sinn

  2. Viral Vector Core Facility, University of Iowa, Iowa City, IA, USA

    Patrick L. Sinn, Jeremy E. Coffin & Kathleen H. Holt

  3. Department of Microbiology, University of Iowa, Iowa City, IA, USA

    Natarajan Ayithan & Wendy Maury

Authors
  1. Patrick L. Sinn
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  2. Jeremy E. Coffin
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  3. Natarajan Ayithan
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  4. Kathleen H. Holt
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  5. Wendy Maury
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Correspondence to Wendy Maury .

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Editors and Affiliations

  1. Friedrich-Loeffler-Institut, Greifswald - Insel Riems, Germany

    Thomas Hoenen

  2. Friedrich-Loeffler-Institut, Greifswald - Insel Riems, Germany

    Allison Groseth

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Sinn, P.L., Coffin, J.E., Ayithan, N., Holt, K.H., Maury, W. (2017). Lentiviral Vectors Pseudotyped with Filoviral Glycoproteins. In: Hoenen, T., Groseth, A. (eds) Ebolaviruses. Methods in Molecular Biology, vol 1628. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7116-9_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7116-9_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7115-2

  • Online ISBN: 978-1-4939-7116-9

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