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 A.
N1 - Funding Information:
The authors thank Theodore Wyman, Kirsten Pilcher, and Tarsem Moudgil for their participation during early phases of this project, and Gail Marracci for the original provision of the D2/RHE/OR/V1 molecular clone used to develop the SRV transduction and packaging cell vectors. We also acknowledge the help of Yibing Jia and Kalama Taylor of the ORPRC Molecular Biology and Cell Culture Core Facilities, for their assistance in automated DNA sequencing and cell culture propagation. We thank Philbert Kirigiti for his assistance in the construction of some of the figures displayed in this manuscript. We also thank other current members of the Machida Laboratory, including Ying Bai and Luz Pell, for encouragement and support. B. Li was a 1997 Leukemia Research Foundation Postdoctoral Fellow, and is currently at the Center for Human Molecular Genetics, Munroe–Meyer Institute, University of Nebraska Medical College. S. Nguyen is now at the Shriner's Hospital for Crippled Children, Portland, Oregon. X. Li is now at the Department of Medical and Molecular Genetics, Oregon Health Sciences University. This research has been supported by grants awarded to C.A. Machida from NIH DK53462, NCRR 00163, the Medical Research Foundation of Oregon, and the Collins Medical Trust.
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 - SRV-based gene transfer system
KW - Simian retrovirus (SRV) vectors
KW - Split-genome packaging cell recombinants
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
C2 - 11325470
AN - SCOPUS:0035014078
SN - 0168-1702
VL - 75
SP - 155
EP - 168
JO - Virus Research
JF - Virus Research
IS - 2
ER -