Recombinant replication-restricted VSV as an expression vector for murine cytokines

Mark Miller, Christy L. Lavine, Sheri D. Klas, Lawrence Pfeffer, Michael Whitt

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Vesicular stomatitis virus (VSV) is a prototypic non-segmented, negative-strand RNA virus that rapidly and efficiently shuts down the production of host cell-encoded proteins and utilizes the cell's protein production machinery to express high levels of virally encoded proteins. In an effort to take advantage of this characteristic of VSV, we have employed a reverse genetics system to create recombinant forms of VSV encoding a variety of murine cytokines. Previous studies have revealed that cells infected with recombinant VSV that lack expression of the surface glycoprotein (G protein), designated ΔG-VSV, more efficiently express and secrete recombinant proteins than do recombinant "wild-type" VSV. Therefore, murine cytokine-expressing recombinants were produced as ΔG viruses. Propagation of these ΔG viruses in cells that transiently express G protein in vitro results in G-complemented virions that can infect cells, shut down host protein synthesis, and express at high levels each virally encoded protein (including the designated cytokine). We assessed the ability of each ΔG-VSV construct to express recombinant cytokine by infecting BHK cells and then monitoring/measuring the production of the desired cytokine. When possible, the bioactivity of the cytokine products was also measured. The results presented here reveal that large quantities of bioactive cytokines can be produced rapidly and inexpensively using ΔG-VSV as a protein expression system.

Original languageEnglish (US)
Pages (from-to)92-103
Number of pages12
JournalProtein Expression and Purification
Volume33
Issue number1
DOIs
StatePublished - Jan 1 2004

Fingerprint

Vesicular Stomatitis
Cytokines
Viruses
Proteins
Reverse Genetics
RNA Viruses
Membrane Glycoproteins
Recombinant Proteins
Virion
Glycoproteins

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Recombinant replication-restricted VSV as an expression vector for murine cytokines. / Miller, Mark; Lavine, Christy L.; Klas, Sheri D.; Pfeffer, Lawrence; Whitt, Michael.

In: Protein Expression and Purification, Vol. 33, No. 1, 01.01.2004, p. 92-103.

Research output: Contribution to journalArticle

@article{6b04f11acf7043239e5e29b3e3ee2515,
title = "Recombinant replication-restricted VSV as an expression vector for murine cytokines",
abstract = "Vesicular stomatitis virus (VSV) is a prototypic non-segmented, negative-strand RNA virus that rapidly and efficiently shuts down the production of host cell-encoded proteins and utilizes the cell's protein production machinery to express high levels of virally encoded proteins. In an effort to take advantage of this characteristic of VSV, we have employed a reverse genetics system to create recombinant forms of VSV encoding a variety of murine cytokines. Previous studies have revealed that cells infected with recombinant VSV that lack expression of the surface glycoprotein (G protein), designated ΔG-VSV, more efficiently express and secrete recombinant proteins than do recombinant {"}wild-type{"} VSV. Therefore, murine cytokine-expressing recombinants were produced as ΔG viruses. Propagation of these ΔG viruses in cells that transiently express G protein in vitro results in G-complemented virions that can infect cells, shut down host protein synthesis, and express at high levels each virally encoded protein (including the designated cytokine). We assessed the ability of each ΔG-VSV construct to express recombinant cytokine by infecting BHK cells and then monitoring/measuring the production of the desired cytokine. When possible, the bioactivity of the cytokine products was also measured. The results presented here reveal that large quantities of bioactive cytokines can be produced rapidly and inexpensively using ΔG-VSV as a protein expression system.",
author = "Mark Miller and Lavine, {Christy L.} and Klas, {Sheri D.} and Lawrence Pfeffer and Michael Whitt",
year = "2004",
month = "1",
day = "1",
doi = "10.1016/j.pep.2003.08.008",
language = "English (US)",
volume = "33",
pages = "92--103",
journal = "Protein Expression and Purification",
issn = "1046-5928",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Recombinant replication-restricted VSV as an expression vector for murine cytokines

AU - Miller, Mark

AU - Lavine, Christy L.

AU - Klas, Sheri D.

AU - Pfeffer, Lawrence

AU - Whitt, Michael

PY - 2004/1/1

Y1 - 2004/1/1

N2 - Vesicular stomatitis virus (VSV) is a prototypic non-segmented, negative-strand RNA virus that rapidly and efficiently shuts down the production of host cell-encoded proteins and utilizes the cell's protein production machinery to express high levels of virally encoded proteins. In an effort to take advantage of this characteristic of VSV, we have employed a reverse genetics system to create recombinant forms of VSV encoding a variety of murine cytokines. Previous studies have revealed that cells infected with recombinant VSV that lack expression of the surface glycoprotein (G protein), designated ΔG-VSV, more efficiently express and secrete recombinant proteins than do recombinant "wild-type" VSV. Therefore, murine cytokine-expressing recombinants were produced as ΔG viruses. Propagation of these ΔG viruses in cells that transiently express G protein in vitro results in G-complemented virions that can infect cells, shut down host protein synthesis, and express at high levels each virally encoded protein (including the designated cytokine). We assessed the ability of each ΔG-VSV construct to express recombinant cytokine by infecting BHK cells and then monitoring/measuring the production of the desired cytokine. When possible, the bioactivity of the cytokine products was also measured. The results presented here reveal that large quantities of bioactive cytokines can be produced rapidly and inexpensively using ΔG-VSV as a protein expression system.

AB - Vesicular stomatitis virus (VSV) is a prototypic non-segmented, negative-strand RNA virus that rapidly and efficiently shuts down the production of host cell-encoded proteins and utilizes the cell's protein production machinery to express high levels of virally encoded proteins. In an effort to take advantage of this characteristic of VSV, we have employed a reverse genetics system to create recombinant forms of VSV encoding a variety of murine cytokines. Previous studies have revealed that cells infected with recombinant VSV that lack expression of the surface glycoprotein (G protein), designated ΔG-VSV, more efficiently express and secrete recombinant proteins than do recombinant "wild-type" VSV. Therefore, murine cytokine-expressing recombinants were produced as ΔG viruses. Propagation of these ΔG viruses in cells that transiently express G protein in vitro results in G-complemented virions that can infect cells, shut down host protein synthesis, and express at high levels each virally encoded protein (including the designated cytokine). We assessed the ability of each ΔG-VSV construct to express recombinant cytokine by infecting BHK cells and then monitoring/measuring the production of the desired cytokine. When possible, the bioactivity of the cytokine products was also measured. The results presented here reveal that large quantities of bioactive cytokines can be produced rapidly and inexpensively using ΔG-VSV as a protein expression system.

UR - http://www.scopus.com/inward/record.url?scp=0347597097&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0347597097&partnerID=8YFLogxK

U2 - 10.1016/j.pep.2003.08.008

DO - 10.1016/j.pep.2003.08.008

M3 - Article

VL - 33

SP - 92

EP - 103

JO - Protein Expression and Purification

JF - Protein Expression and Purification

SN - 1046-5928

IS - 1

ER -