The intent is to design retrovirus vectors to a) study retrovirus replication, b) optimize the efficiency and usefulness of gene transfer, and c) study cis-acting elements that regulate gene expression. The principle vector will be a derivative of mouse amphotropic virus 4070 to maximize host range and allow efficient gene transfer and expression in human cells. The system will be designed to eliminate the need for a helper virsus or phenotypic selection and will not require the synthesis of any viral proteins or RNA in the target cells. The methodologies involved include recombinant DNA cloning, in vitro mutagenesis of DNA, growth of virus in tissue culture, and analysis of gene expression using gel electrophoresis and nucleic acid hybridization. The long term goals are a) to better understand retrovirus replication and pathogenesis, b) to understand the regulation of gene expression, and c) to develop a gene transfer system that will be useful in both somatic cell and germline-directed gene transfer. The health-relatedness of the project will stem from its potential usage in gene therapy. A more complete understanding of retrovirus replication and expression will increase our understanding of oncogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA036448-03
Application #
3174029
Study Section
Experimental Virology Study Section (EVR)
Project Start
1983-12-01
Project End
1986-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
San Diego
State
CA
Country
United States
Zip Code
92037
Mullinax, R L; Gross, E A; Amberg, J R et al. (1990) Identification of human antibody fragment clones specific for tetanus toxoid in a bacteriophage lambda immunoexpression library. Proc Natl Acad Sci U S A 87:8095-9
Flamant, F; Sorge, J A (1988) In vitro synthesis of infectious retroviral RNA. J Virol 62:1827-31
Sorge, J A; West, C; Kuhl, W et al. (1987) The human glucocerebrosidase gene has two functional ATG initiator codons. Am J Hum Genet 41:1016-24
Flamant, F; Gurin, C C; Sorge, J A (1987) An embryonic DNA-binding protein specific for the promoter of the retrovirus long terminal repeat. Mol Cell Biol 7:3548-53
Sorge, J; Kuhl, W; West, C et al. (1987) Complete correction of the enzymatic defect of type I Gaucher disease fibroblasts by retroviral-mediated gene transfer. Proc Natl Acad Sci U S A 84:906-9
Jirik, F R; Burstein, S A; Treger, L et al. (1987) Transfection of a factor-dependent cell line with the murine interleukin-3 (IL-3) cDNA results in autonomous growth and tumorigenesis. Leuk Res 11:1127-34
Radoux, V; Chen, P P; Sorge, J A et al. (1986) A conserved human germline V kappa gene directly encodes rheumatoid factor light chains. J Exp Med 164:2119-24
Sorge, J; Kuhl, W; West, C et al. (1986) Gaucher disease: retrovirus-mediated correction of the enzymatic defect in cultured cells. Cold Spring Harb Symp Quant Biol 51 Pt 2:1041-6
Sorge, J; West, C; Westwood, B et al. (1985) Molecular cloning and nucleotide sequence of human glucocerebrosidase cDNA. Proc Natl Acad Sci U S A 82:7289-93
Sorge, J; Gelbart, T; West, C et al. (1985) Heterogeneity in type I Gaucher disease demonstrated by restriction mapping of the gene. Proc Natl Acad Sci U S A 82:5442-5