Simian retrovirus vectors for gene transfer in nonhuman primate cells

Biao Li, Susan Nguyen, Xiaorong Li, Curtis Machida

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We have recently identified and sequenced a molecular clone of the serogroup 2 simian retrovirus (SRV), D2/RHE/OR/V1, that retains an enhanced ability to infect specific T cell lines. In this report, using deletion mutagenesis, we localized the psi packaging signal, necessary for packaging of D2/RHE/OR/V1 particles, to the genomic region 345-650, which comprises the 5′ intergenic region (IR) and the extreme 5′ portion of the gag gene. To build an SRV-based gene transfer system and to reduce the possibility of recombination and regeneration of replication-competent viruses, we constructed split-genome D2/RHE/OR/V1 plasmid recombinants containing distinct and non-overlapping retroviral gene regions and several replacement components. For the retrovirus gene transfer vehicle, we deleted the D2/RHE/OR/V1 structural genes and substituted a cassette including the psi-packaging region, the β-galactosidase reporter gene, and the 3′ IR. Both packaging cell recombinants were used to generate stable monkey packaging cell lines; the gene transfer vehicle was subsequently transfected into the packaging cell lines, and replication-defective viruses were recovered for subsequent infection into fresh monkey cells. Successful infection by the recovered viruses verifies the potential efficacy of the SRV-based system as a research tool for gene transfer of heterologous genes into nonhuman primate cells.

Original languageEnglish (US)
Pages (from-to)155-168
Number of pages14
JournalVirus Research
Volume75
Issue number2
DOIs
StatePublished - Jun 2001

Fingerprint

Simian Retroviruses
Primates
Product Packaging
Genes
Intergenic DNA
Cell Line
Haplorhini
Mason-Pfizer monkey virus
Galactosidases
gag Genes
Defective Viruses
Virus Diseases
Retroviridae
Virus Replication
Reporter Genes
Mutagenesis
Genetic Recombination
Regeneration
Plasmids
Clone Cells

Keywords

  • D2/RHE/OR/V1 serogroup 2 SRV
  • Psi packaging signal
  • Simian retrovirus (SRV) vectors
  • Split-genome packaging cell recombinants
  • SRV-based gene transfer system

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Simian retrovirus vectors for gene transfer in nonhuman primate cells. / Li, Biao; Nguyen, Susan; Li, Xiaorong; Machida, Curtis.

In: Virus Research, Vol. 75, No. 2, 06.2001, p. 155-168.

Research output: Contribution to journalArticle

Li, Biao ; Nguyen, Susan ; Li, Xiaorong ; Machida, Curtis. / Simian retrovirus vectors for gene transfer in nonhuman primate cells. In: Virus Research. 2001 ; Vol. 75, No. 2. pp. 155-168.
@article{b7dc651ba2bc44b1816c802ccdfc8345,
title = "Simian retrovirus vectors for gene transfer in nonhuman primate cells",
abstract = "We have recently identified and sequenced a molecular clone of the serogroup 2 simian retrovirus (SRV), D2/RHE/OR/V1, that retains an enhanced ability to infect specific T cell lines. In this report, using deletion mutagenesis, we localized the psi packaging signal, necessary for packaging of D2/RHE/OR/V1 particles, to the genomic region 345-650, which comprises the 5′ intergenic region (IR) and the extreme 5′ portion of the gag gene. To build an SRV-based gene transfer system and to reduce the possibility of recombination and regeneration of replication-competent viruses, we constructed split-genome D2/RHE/OR/V1 plasmid recombinants containing distinct and non-overlapping retroviral gene regions and several replacement components. For the retrovirus gene transfer vehicle, we deleted the D2/RHE/OR/V1 structural genes and substituted a cassette including the psi-packaging region, the β-galactosidase reporter gene, and the 3′ IR. Both packaging cell recombinants were used to generate stable monkey packaging cell lines; the gene transfer vehicle was subsequently transfected into the packaging cell lines, and replication-defective viruses were recovered for subsequent infection into fresh monkey cells. Successful infection by the recovered viruses verifies the potential efficacy of the SRV-based system as a research tool for gene transfer of heterologous genes into nonhuman primate cells.",
keywords = "D2/RHE/OR/V1 serogroup 2 SRV, Psi packaging signal, Simian retrovirus (SRV) vectors, Split-genome packaging cell recombinants, SRV-based gene transfer system",
author = "Biao Li and Susan Nguyen and Xiaorong Li and Curtis Machida",
year = "2001",
month = "6",
doi = "10.1016/S0168-1702(01)00236-2",
language = "English (US)",
volume = "75",
pages = "155--168",
journal = "Virus Research",
issn = "0168-1702",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Simian retrovirus vectors for gene transfer in nonhuman primate cells

AU - Li, Biao

AU - Nguyen, Susan

AU - Li, Xiaorong

AU - Machida, Curtis

PY - 2001/6

Y1 - 2001/6

N2 - We have recently identified and sequenced a molecular clone of the serogroup 2 simian retrovirus (SRV), D2/RHE/OR/V1, that retains an enhanced ability to infect specific T cell lines. In this report, using deletion mutagenesis, we localized the psi packaging signal, necessary for packaging of D2/RHE/OR/V1 particles, to the genomic region 345-650, which comprises the 5′ intergenic region (IR) and the extreme 5′ portion of the gag gene. To build an SRV-based gene transfer system and to reduce the possibility of recombination and regeneration of replication-competent viruses, we constructed split-genome D2/RHE/OR/V1 plasmid recombinants containing distinct and non-overlapping retroviral gene regions and several replacement components. For the retrovirus gene transfer vehicle, we deleted the D2/RHE/OR/V1 structural genes and substituted a cassette including the psi-packaging region, the β-galactosidase reporter gene, and the 3′ IR. Both packaging cell recombinants were used to generate stable monkey packaging cell lines; the gene transfer vehicle was subsequently transfected into the packaging cell lines, and replication-defective viruses were recovered for subsequent infection into fresh monkey cells. Successful infection by the recovered viruses verifies the potential efficacy of the SRV-based system as a research tool for gene transfer of heterologous genes into nonhuman primate cells.

AB - We have recently identified and sequenced a molecular clone of the serogroup 2 simian retrovirus (SRV), D2/RHE/OR/V1, that retains an enhanced ability to infect specific T cell lines. In this report, using deletion mutagenesis, we localized the psi packaging signal, necessary for packaging of D2/RHE/OR/V1 particles, to the genomic region 345-650, which comprises the 5′ intergenic region (IR) and the extreme 5′ portion of the gag gene. To build an SRV-based gene transfer system and to reduce the possibility of recombination and regeneration of replication-competent viruses, we constructed split-genome D2/RHE/OR/V1 plasmid recombinants containing distinct and non-overlapping retroviral gene regions and several replacement components. For the retrovirus gene transfer vehicle, we deleted the D2/RHE/OR/V1 structural genes and substituted a cassette including the psi-packaging region, the β-galactosidase reporter gene, and the 3′ IR. Both packaging cell recombinants were used to generate stable monkey packaging cell lines; the gene transfer vehicle was subsequently transfected into the packaging cell lines, and replication-defective viruses were recovered for subsequent infection into fresh monkey cells. Successful infection by the recovered viruses verifies the potential efficacy of the SRV-based system as a research tool for gene transfer of heterologous genes into nonhuman primate cells.

KW - D2/RHE/OR/V1 serogroup 2 SRV

KW - Psi packaging signal

KW - Simian retrovirus (SRV) vectors

KW - Split-genome packaging cell recombinants

KW - SRV-based gene transfer system

UR - http://www.scopus.com/inward/record.url?scp=0035014078&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035014078&partnerID=8YFLogxK

U2 - 10.1016/S0168-1702(01)00236-2

DO - 10.1016/S0168-1702(01)00236-2

M3 - Article

VL - 75

SP - 155

EP - 168

JO - Virus Research

JF - Virus Research

SN - 0168-1702

IS - 2

ER -