Long-term persistence of plasmid DNA and foreign gone expression in mouse muscle
| Autor: | Jon A. Wolff, Philip Williams, Gyula Acsadi, James J. Ludtke, Agnes Jani |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Chloramphenicol O-Acetyltransferase
Time Factors Gene Expression Biology Transfection Polymerase Chain Reaction Cell Line Mice chemistry.chemical_compound Plasmid Extrachromosomal DNA Genetics medicine Animals Luciferases Molecular Biology Genetics (clinical) Muscles Electroporation Skeletal muscle DNA General Medicine Molecular biology Real-time polymerase chain reaction medicine.anatomical_structure chemistry Ligation Plasmids |
| Zdroj: | Human Molecular Genetics. 1:363-369 |
| ISSN: | 1460-2083 0964-6906 |
| DOI: | 10.1093/hmg/1.6.363 |
| Popis: | Plasmid pRSVL persisted and expressed luciferase for at least 19 months in mouse skeletal muscle after intramuscular injection. Other injected plasmids also stably expressed long-term suggesting that any plasmid DNA could stably persist and express in muscle. Plasmid DNA was demonstrated by quantitative PCR in some of the muscle DNA samples for at least 19 months after injection. The methylation pattern of the plasmid DNA remained in its bacterial form indicating that the foreign DNA did not replicate in the muscle cells. The electroporation of total cellular DNA from injected muscles into bacteria indicated that the plasmid DNA was extrachromosomal. Chromosomal integration of plasmid DNA was searched for by electroporating the injected muscle DNA into bacteria after restriction enzyme digestion and ligation. No plasmids containing plasmid/chromosome junctions were observed in over 1800 colonies examined. Lack of integration increases the theoretical safety of this gene transfer technique. Long-term stability of plasmid DNA in muscle indicates that muscle is an attractive target tissue for the introduction of extrachromosomal plasmid or viral DNA for the purpose of gene therapy. |
| Databáze: | OpenAIRE |
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