Long-Term Expression of a Fluorescent Reporter Gene via Direct Injection of Plasmid Vector into Mouse Skeletal Muscle: Comparison of Human Creatine Kinase and Cmv Promoter Expression Levels in Vivo
| Autor: | R J, Bartlett, S L, Secore, J T, Singer, M, Bodo, K, Sharma, C, Ricordi |
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| Rok vydání: | 1996 |
| Předmět: |
0301 basic medicine
Cell Transplantation Genetic Vectors Green Fluorescent Proteins Molecular Sequence Data Restriction Mapping Biomedical Engineering Cytomegalovirus lcsh:Medicine Transfection Polymerase Chain Reaction Mice 03 medical and health sciences 0302 clinical medicine Genes Reporter Animals Humans Muscle Skeletal Promoter Regions Genetic Creatine Kinase DNA Primers Transplantation Base Sequence lcsh:R Cell Biology Isoenzymes Luminescent Proteins 030104 developmental biology Mice Inbred mdx 030217 neurology & neurosurgery Plasmids |
| Zdroj: | Cell Transplantation, Vol 5 (1996) |
| ISSN: | 1555-3892 0963-6897 |
| DOI: | 10.1177/096368979600500308 |
| Popis: | Expression of a fluorescent reporter gene has been studied using two alternate promoters to transcribe the green fluorescent protein (gfp) from Aequorea victoria. The human cytomegalovirus (CMV) enhancer/promoter or the human muscle-specific creatine kinase promoter (CKM) were inserted along with the gfp cDNA into a plasmid expression vector based on a modified adeno-associated virus genome. Naked plasmid DNA was injected into the hamstring muscle of mdx mice and gfp gene expression determined from frozen muscle sections taken at 4, 14, and 42 days postinjection. Fluorescence patterns obtained by photomicroscopy and quantitative fluorescence measurements indicated a near-linear increase in the accumulation of the gfp in skeletal muscle during the length of the study, with gfp expression at 42 days being roughly four times the values obtained at 4 days. The levels of expression of gfp from the CKM construct were consistantly higher than for the CMV construct. The CKM promoter/expression vector combination demonstrates significant potential for simple, direct delivery and long-term, high-level expression of genes in skeletal muscle. |
| Databáze: | OpenAIRE |
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