The role of plasmid constructs containing the SV40 DNA nuclear-targeting sequence in cationic lipid-mediated DNA delivery.
One of the steps that limit transfection efficiency in non-viral gene delivery is inefficient nuclear import of plasmid DNA, once it has been delivered into the cytoplasm. Recently, via microinjection into the cytoplasm and in situ hybridizations into a few cell types, it was shown that a region of Simian virus 40(SV40), specifically a c. 372-bp fragment of SV40 genomic DNA encompassing the SV40 promoter-enhancer-origin of replication (SV40 DTS), could enable the nuclear import of a plasmid carrying these sequences (Dean D.A. Exp. Cell Res. 230 (1997) 293). In this report, we address the issue of the suitability of the SV40 DTS for cationic lipid-mediated gene delivery, and its capacity to improve the efficiency of the transfection process. For this study, we used transient reporter gene expression assays on various cell types. The gene expression from the plasmid constructs carrying the SV40 DTS varied with cell type and plasmid construct used. Such cell-type and plasmid-construct dependency on gene expression from plasmids containing the SV40 DTS suggests that the gene expression from plasmids is not entirely dependent on its ability to enhance the nuclear import of said plasmids.
Prasad
Tekkatte Krishnamurthy
TK
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad (AP), India 500007.
Rao
Nalam Madhusudhana
NM
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D001692:Biological Transport; D016466:CHO Cells; D019556:COS Cells; D002412:Cations; D002467:Cell Nucleus / Q000378:metabolism*; D006224:Cricetinae; D004261:DNA Replication / Q000187:drug effects; D005434:Flow Cytometry; D015870:Gene Expression; D017930:Genes, Reporter; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D010957:Plasmids / Q000235:genetics*; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection / Q000295:instrumentation / Q000379:methods*
D002412:Cations; D008055:Lipids; D008081:Liposomes; D016590:Aphidicolin