TRIPLE HYBRID AMPLICON VECTOR SYSTEMS TO GENERATE RETROVIRAL PACKAGING LINES

摘要

The present invention relates to a triple hybrid vector amplicon system comprising genetic elements derived from Herpes Simplex Virus (HSV), Epstein-Barr Virus (EBV) or Adeno-Associated Virus (AAV), and a retrovirus. The vector was developed to stably transform cells, both in culture or in vivo, into retrovirus packaging cells in a single step. This step can be accomplished both by transfection using liposomes, electroporation, calcium phosphate, or any other methodology used to transfer naked or complexed DNA into cells or by infection when the vector is packaged as an amplicon vector in HSV virions. In one embodiment, the system takes advantage of the host range and retention properties of HSV/EBV hybrid amplicons to efficiently convert cells to retrovirus vector producer cells after single-step transduction. Retrovirus genes gag-pol and env (GPE) and retroviral vector sequences were modified to minimize sequence overlap and cloned into an HSV/EBV hybrid amplicon. In a second embodiment, retrovirus gag-pol and env genes and a retrovirus vector carrying lacZ, were cloned into HSV/AAV hybrid amplicons. These hybrid amplicon vector systems hold great potential for the generation of new retrovirus packaging lines derived from cells that due to their migratory, tumor or tissue targeting properties, can expand the spatial distribution of gene delivery by retrovirus vectors in vivo.