Efficient gene transfer into the CNS by lentiviral vectors purified by anion exchange chromatography
| Autor: | S Schüle, Michaela Scherr, Matthias Eder, Arnold Ganser, U Blömer, Manuel Grez, Karin Battmer, H Hohenberg |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Transgene
Genetic enhancement Genetic Vectors Green Fluorescent Proteins Biology Viral vector Transduction (genetics) In vivo Transduction Genetic Genetics Animals Vector (molecular biology) Molecular Biology Cells Cultured Neurons Chromatography Genetic transfer Lentivirus Brain Chromatography Ion Exchange In vitro Rats Inbred F344 Rats Luminescent Proteins Molecular Medicine Female Neuroglia |
| Zdroj: | Gene therapy. 9(24) |
| ISSN: | 0969-7128 |
| Popis: | Lentiviral vectors have been shown to stably transduce dividing and non-dividing target cells in vitro and in vivo. However, in vivo gene transfer applications with viral vectors in the central nervous system require highly efficient vector preparations, because only very small volumes can be injected stereotactically without damage to the brain tissue. Since lentiviral vectors are generated in transient co-transfection systems, viral preparations need to be purified and efficiently concentrated before injection into the brain. We describe an alternative procedure to concentrate lentiviral preparations by binding viral particles to an anion exchange column. Viral particles are eluted with sodium chloride, desalted and further concentrated by ultrafiltration. These vector preparations allowed high levels of gene transfer into terminally differentiated neuronal and glial cells and long-term transgene expression without any signs of acute and long-term toxicity or inflammation. The purification of lentiviral vectors from large-scale preparations by anion exchange chromatography allowed us to concentrate the virus to small volumes and to use these preparations to genetically modified target cells in vivo without signs of acute inflammatory responses. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|