The destabilization of polyplexes facilitates intranuclear transcription efficiency.
We have been focusing on the last step of transfection, that is, the intranuclear recognition of polyplexes by transcription factors. In this work, we studied about the relationship between the intracellular destabilization of polyplexes and the transcription efficiency. Polypeptides containing specific sequences which are digested by intracellular proprotein convertase (PC), furin (Fur-polypeptide) were synthesized as novel carriers. When Fur-polypeptide/plasmid DNA polyplexes were incubated with furin in the cell-free transcription/translation system, the sequence specific expression was observed as time. Additionally, Fur-polypeptides/plasmid DNA polyplexes led to site-specific effective transgene expression even in COS-1 cells. These results indicate that shortened carriers lose their abilities to form polyplexes in response to furin activity in the cells, and it seems to enhance the destabilization of polyplexes, the recognition of transgene in polyplexes by transcription factors, and the expression efficiency.
Hashimoto
Tomoko
T
Department of Biomedical Engineering, National Cardiovascular Center Research Institute, Fujishirodai, Suita, Osaka 565-8585, Japan.
Kobori
Akio
A
Murakami
Akira
A
Yamaoka
Tetsuji
T
D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000235:genetics; D002522:Chlorocebus aethiops; D004247:DNA / Q000008:administration & dosage; D045683:Furin / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011107:Polylysine / Q000737:chemistry; D014158:Transcription, Genetic; D014162:Transfection; D019076:Transgenes
D010455:Peptides; D011107:Polylysine; D004247:DNA; D045683:Furin