Efficient in vivo gene transfer by PCR amplified fragment with reduced inflammatory activity View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2001-01

AUTHORS

CR Hofman, JP Dileo, Z Li, S Li, L Huang

ABSTRACT

There are many problems associated with plasmid DNA that may limit its use in systemic gene transfer. These problems could be solved by the use of synthetic genes. As a model to test the feasibility of using synthetic genes for gene therapy, we PCR-amplified a fragment containing the CMV promoter, the luciferase gene and a polyadenylation signal. The in vivo expression efficiency of the PCR fragment was determined by using two different methods, a hydrodynamics-based gene transfer of naked DNA to the liver and LPD (a lipid-based vector) mediated gene transfer to the lung. Our results show that linear fragments are at least as active as plasmid DNA following systemic delivery by LPD. However, PCR fragments are much less inflammatory than plasmid DNA as shown by a three-fold reduction in serum levels of both TNF-alpha and IL-12. Our results also showed that PCR fragments are highly efficient in liver gene transfer following systemic administration in a large volume. Thus, these results support the idea of using synthetic genes for gene therapy. Since gene sequence can be easily obtained as a PCR fragment, our results also imply that it may provide a useful and convenient method for determining the physiologic function of a putative gene in intact animals. More... »

PAGES

71

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/sj.gt.3301373

DOI

http://dx.doi.org/10.1038/sj.gt.3301373

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1017095695

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/11402304


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