A single-plasmid reverse genetics system for the rescue of non-segmented negative-strand RNA viruses from cloned full-length cDNA

Ben Peeters*, Olav de Leeuw

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

Reverse genetics systems for non-segmented negative-strand RNA viruses rely on co-transfection of a plasmid containing the full-length viral cDNA and helper plasmids encoding essential viral replication proteins. Here, a system is presented in which virus can be rescued from a single plasmid without the need for helper plasmids in cells infected with a host-restricted recombinant poxvirus that expresses T7 RNA polymerase. This approach relies on the insertion of T7 promoter sequences in the viral cDNA at positions that allow transcription of sub-genomic RNAs encoding essential viral replication proteins.

Original languageEnglish
Pages (from-to)187-190
JournalJournal of Virological Methods
Volume248
DOIs
Publication statusPublished - 2017

Fingerprint

Reverse Genetics
RNA Viruses
Plasmids
Complementary DNA
Viral Proteins
Poxviridae
Transfection
RNA
Viruses

Keywords

  • Negative-strand RNA virus
  • Reverse genetics
  • T7-promoter

Cite this

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abstract = "Reverse genetics systems for non-segmented negative-strand RNA viruses rely on co-transfection of a plasmid containing the full-length viral cDNA and helper plasmids encoding essential viral replication proteins. Here, a system is presented in which virus can be rescued from a single plasmid without the need for helper plasmids in cells infected with a host-restricted recombinant poxvirus that expresses T7 RNA polymerase. This approach relies on the insertion of T7 promoter sequences in the viral cDNA at positions that allow transcription of sub-genomic RNAs encoding essential viral replication proteins.",
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A single-plasmid reverse genetics system for the rescue of non-segmented negative-strand RNA viruses from cloned full-length cDNA. / Peeters, Ben; de Leeuw, Olav.

In: Journal of Virological Methods, Vol. 248, 2017, p. 187-190.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - A single-plasmid reverse genetics system for the rescue of non-segmented negative-strand RNA viruses from cloned full-length cDNA

AU - Peeters, Ben

AU - de Leeuw, Olav

PY - 2017

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N2 - Reverse genetics systems for non-segmented negative-strand RNA viruses rely on co-transfection of a plasmid containing the full-length viral cDNA and helper plasmids encoding essential viral replication proteins. Here, a system is presented in which virus can be rescued from a single plasmid without the need for helper plasmids in cells infected with a host-restricted recombinant poxvirus that expresses T7 RNA polymerase. This approach relies on the insertion of T7 promoter sequences in the viral cDNA at positions that allow transcription of sub-genomic RNAs encoding essential viral replication proteins.

AB - Reverse genetics systems for non-segmented negative-strand RNA viruses rely on co-transfection of a plasmid containing the full-length viral cDNA and helper plasmids encoding essential viral replication proteins. Here, a system is presented in which virus can be rescued from a single plasmid without the need for helper plasmids in cells infected with a host-restricted recombinant poxvirus that expresses T7 RNA polymerase. This approach relies on the insertion of T7 promoter sequences in the viral cDNA at positions that allow transcription of sub-genomic RNAs encoding essential viral replication proteins.

KW - Negative-strand RNA virus

KW - Reverse genetics

KW - T7-promoter

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DO - 10.1016/j.jviromet.2017.07.008

M3 - Article

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JO - Journal of Virological Methods

JF - Journal of Virological Methods

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