High mutation rate of a spleen necrosis virus-based retrovirus vector.
Spleen necrosis virus (SNV) is an avian retrovirus that efficiently infects some mammalian cells (e.g., dog and rat cells). We constructed an SNV-based vector, which contains less than 1 kilobase (kb) of the retrovirus sequence, and a number of derivatives containing selectable markers. We obtained high-titer virus stocks, over 10(6) transforming units per ml, with a vector whose genomic RNA consists of 1,850 bases (full-length SNV RNA is 7.7 kb). We also studied two vectors that both carry two genes which should be expressed from a single promoter, one gene from unspliced mRNA and the other gene from spliced mRNA. In one vector, both genes were efficiently expressed as expected. However, in the other vector, expression of the gene 3' to the splice acceptor was inhibited. When we selected for expression of the 3' gene is this latter case, we found that the resistant cells contained mutant proviruses in which the 3' gene could be expressed. Furthermore, we found that mutations were generated during a single round of virus replication (provirus to provirus) at a rate of approximately 0.5% mutations per cycle.
D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D002642:Chick Embryo; D002999:Clone Cells; D004285:Dogs; D005347:Fibroblasts; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection