by Jurgen Seppen, Simon C. Barry, Brandon Harder, and William R. A

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Lentivirus administration to rat muscle provides efficient sustained expression of erythropoietin by Jurgen Seppen, Simon C. Barry, Brandon Harder, and William R. A. Osborne Blood Volume 98(3):594-596 August 1, 2001 ©2001 by American Society of Hematology

Serial analysis of percent hematocrit of EPO-lentivirus–treated and control rats.Closed symbols are rats receiving EPO-lentivirus 6 × 107IU (●) or 6 × 106 IU (▴). Serial analysis of percent hematocrit of EPO-lentivirus–treated and control rats.Closed symbols are rats receiving EPO-lentivirus 6 × 107IU (●) or 6 × 106 IU (▴). Open symbols are control rats receiving saline (○), eGFP lentivirus 5 × 106 IU (■) or 100 μg lentivirus plasmid (▵). Jurgen Seppen et al. Blood 2001;98:594-596 ©2001 by American Society of Hematology

PCR assay of provirus.Genomic DNA was isolated from tissue samples and subjected to PCR using EPO-specific primers. PCR assay of provirus.Genomic DNA was isolated from tissue samples and subjected to PCR using EPO-specific primers. PCR products were separated on a 2% agarose gel and stained with ethidium bromide. Shown is a PCR on negative control rat DNA in which increasing amounts of vector plasmid DNA were added. We established that the presence of 1% vector DNA could be detected by this method. Shown are assays of representative tissue DNA samples from a rat with an hematocrit of 74% killed 14 months after injection of EPO virus. Lane 1, molecular weight marker; lane 2, genomic DNA; lane 3, genomic DNA + 0.1% vector; lane 4, genomic DNA + 1% vector; lane 5, genomic DNA + 10% vector; lane 6, genomic DNA + 100% vector; lane 7, genomic DNA + 1000% vector; lane 8, vector; lane 9, water; lane 10, lung; lane 11, kidney; lane 12, liver; lane 13, spleen; lanes 14 to 17, muscle samples. Jurgen Seppen et al. Blood 2001;98:594-596 ©2001 by American Society of Hematology