Enhancer and Promoter Chimeras in Plasmids Designed for Intramuscular Injection: A ComparativeIn VivoandIn VitroStudy
| Autor: | Marston Manthorpe, Jukka Hartikka, Jon Norman, Peter Hobart, KM Barnhart |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Cytomegalovirus
Gene Expression Biology Injections Intramuscular Cell Line Mice Plasmid In vivo Cricetinae Genetics Baby hamster kidney cell Animals Luciferases Promoter Regions Genetic Enhancer Genes Immediate-Early Molecular Biology Gene DNA Primers Base Sequence Chimera Promoter Transfection Molecular biology Enhancer Elements Genetic Regulatory sequence Molecular Medicine Plasmids |
| Zdroj: | Human Gene Therapy. 9:2545-2553 |
| ISSN: | 1557-7422 1043-0342 |
| Popis: | Enhancers and promoters from various muscle-specific genes were substituted for or combined with the enhancer/promoter of the human cytomegalovirus (CMV) IE gene in a luciferase reporter gene plasmid in an effort to identify new promoter chimeras with increased expression activity after direct intramuscular injection. The regulatory sequence substitutions or additions varied in content, location, and orientation relative to the CMV regulatory sequences. The expression activities of the derivative and parent plasmids were compared quantitatively in vivo using a standard mouse intramuscular injection assay, and in vitro by transfection of differentiated C2C12 mouse myoblasts and BHK hamster kidney cells, to test whether cultured cell transfection could substitute for at least some animal experimentation. In vivo, 1 of 19 of the enhancer/promoter chimeras increased expression levels. In vitro, some chimeras showed significant expression augmentation in C2C12 cells, but not in BHK cells. We conclude that because of differences in plasmid expression profiles, these cell culture systems cannot readily substitute for in vivo testing of new plasmid constructs. |
| Databáze: | OpenAIRE |
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