Spleen necrosis virus-derived C-type retroviral vectors for gene transfer to quiescent cells.
Gene therapy applications of retroviral vectors derived from C-type retroviruses have been limited to introducing genes into dividing target cells. Here, we report genetically engineered C-type retroviral vectors derived from spleen necrosis virus (SNV), which are capable of infecting nondividing cells. This has been achieved by introducing a nuclear localization signal (NLS) sequence into the matrix protein (MA) of SNV by site-directed mutagenesis. This increased the efficiency of infecting nondividing cells and was sufficient to endow the virus with the capability to efficiently infect growth-arrested human T lymphocytes and quiescent primary monocyte-derived macrophages. We demonstrate that this vector actively penetrates the nucleus of a target cell, and has potential use as a gene therapy vector to transfer genes into nondividing cells.
Parveen
Z
Z
The Dorrance H. Hamilton Laboratories, Thomas Jefferson University, Division of Infectious Diseases, Center for Human Virology, 1020 Locust Street, Suite 329, Philadelphia, PA 19107, USA.
Krupetsky
A
A
Engelstädter
M
M
Cichutek
K
K
Pomerantz
R J
RJ
Dornburg
R
R
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D002455:Cell Division; D002460:Cell Line; D002467:Cell Nucleus / Q000821:virology; D004285:Dogs; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005434:Flow Cytometry; D017988:Gammaretrovirus / Q000378:metabolism*; D015683:Gene Products, gag / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D008264:Macrophages / Q000821:virology; D008969:Molecular Sequence Data; D009000:Monocytes / Q000821:virology; D016297:Mutagenesis, Site-Directed; D009336:Necrosis; D019913:Nuclear Localization Signals / Q000235:genetics; D010957:Plasmids; D013154:Spleen / Q000821:virology; D013601:T-Lymphocytes / Q000821:virology; D014162:Transfection; D014407:Tumor Cells, Cultured
D016326:Extracellular Matrix Proteins; D015683:Gene Products, gag; D019913:Nuclear Localization Signals