Replication-defective (E1+E3 deleted) human adenovirus vectors are promising means of therapeutic gene delivery to skeletal muscle cells. Since the tropism of adenovirus is non-selective, muscle-specific expression of systemically administered vectors can only be achieved by the use of a tissue-specific promoter/enhancer that is small enough to fit the insert capacity of the vector. We have generated a replication-defective adenovirus recombinant (AV) in which the reporter gene (firefly luciferase) was driven by a truncated (1.35 kb) muscle creatine kinase (MCK) promoter/enhancer. Highly efficient and muscle-specific transgene expression was demonstrated in immunodeficient mice after local injection of AV into muscles at early age. Luciferase levels produced by AVMCKlux compared favourably to those in parallel experiments from injection of AVRSVlux in which lux expression is driven by the ubiquitously active LTR sequences of RSV. In nonmuscle tissues (brain, liver, kidney, lung), the transgene expression was extremely low even though in these tissues in situ polymerase chain reaction showed as high an infectivity of the cells by the AV as in muscle, and high levels of expression were obtained with AVRSVlux. The relatively small size, the good efficiency and the muscle specificity of the MCK promoter/enhancer would make it ideal to drive the 6.3 kb (truncated) dystrophin cDNA in first generation AV (with a limited (8 kb) insert capacity) designed for gene therapy of Duchenne muscular dystrophy.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.20231 |
| Date | January 1998 |
| Creators | Larochelle, Nancy. |
| Contributors | Nalbantoglu, Josephine (advisor), Karpati, George (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Neurology and Neurosurgery.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001643468, proquestno: MQ44094, Theses scanned by UMI/ProQuest. |
Page generated in 0.0018 seconds