An XMRV Derived Retroviral Vector as a Tool for Gene Transfer
© Cervantes-Garcia et al; licensee BioMed Central Ltd. 2011
Received: 20 May 2011
Accepted: 8 June 2011
Published: 8 June 2011
Retroviral vectors are widely used tools for gene delivery and gene therapy. They are useful for gene expression studies and genetic manipulation in vitro and in vivo. Many retroviral vectors are derived from the mouse gammaretrovirus, murine leukemia virus (MLV). These vectors have been widely used in gene therapy clinical trials. XMRV, initially found in prostate cancer tissue, was the first human gammaretrovirus described.
We developed a new retroviral vector based on XMRV called pXC. It was developed for gene transfer to human cells and is produced by transient cotransfection of LNCaP cells with pXC and XMRV-packaging plasmids.
We demonstrated that pXC mediates expression of inserted transgenes in cell lines. This new vector will be a useful tool for gene transfer in human and non-human cell lines, including gene therapy studies.
Retroviral vectors offer a highly efficient method of stable gene transfer in mammalian cells due to their ability to integrate into the host genome [1, 2]. Moreover, the common genetic architecture of most retroviruses allows the development of similar retroviral vectors with different potentials for cell entry via virus-specific receptors and different capabilities for gene expression mediated by diverse retroviral promoters .
Current retroviral vectors used for gene transfer are replication defective. Trans-expression of retroviral structural proteins from non vector-homologous plasmids avoids the production of replication competent retrovirus (RCR) [4, 5]. Many retroviral vectors are derived from murine leukemia virus (MLV) in both, ecotropic and amphotropic versions [6, 7]. Lentiviral vectors based on HIV may offer advantages because of their lower insertion frequency in crucial loci involved in cell growth regulation and their ability to transduce non-dividing cells [8, 9]. Nevertheless, MLV-derived retroviral vectors have been used extensively, including in more than 300 gene therapy clinical trials . In addition, retroviral vectors derived from avian sarcoma leukosis virus (ASLV; ), spleen focus-forming virus (SFFV; ), and Mason-Pfizer monkey virus (MPMV; ), have been developed among others.
In 2006, the xenotropic murine-leukemia-virus related gammaretrovirus (XMRV) was discovered in a subset of human prostate cancer (PCa) tissue samples . Subsequently, additional studies demonstrated that XMRV uses the XPR1 receptor to initiate infection, and that the virus is sensitive to IFN-β and RNase-L, a final effector of the IFN-β mediated antiviral response . Since XMRV shows the basic structure of gammaretroviruses, we developed a panel of packaging plasmids and retroviral vectors derived from XMRV. Here we demonstrate their potential use as gene transfer vectors for in vitro assays.
Oligonucleotide primers used in this study.
Luciferase expression plasmid
Packaging plasmid gag-pol
Packaging plasmid env
Many retroviral vectors for gene transfer are derived from murine gamma-retroviruses. Nevertheless, to our knowledge, the pXC vector and its associated cell packaging system described here constitute the first retroviral vector system for gene transfer based on a likely human gammaretrovirus, with particular tropism for human cells and tissues. Our results suggest that the pXC vector is a useful tool for gene transfer in human cells and it is possible that this vector will contribute to elucidation of the interactions of XMRV and its human host. In addition, this new XMRV-derived retroviral vector has the potential to be used for stable transfection of human cells and for preclinical studies of gene therapy.
We thank UC-MEXUS CONACyT for support of this study with grant number CN08-229. We also thank Drs. Sandra and Francis Ruscetti, and Dr. John Elder, for providing primary antibodies.
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