Defective adenoassociated viral-mediated transfection of insulin gene by direct injection into liver parenchyma decreases blood glucose of diabetic mice Journal Article
| Authors: | Sugiyama, A.; Hattori, S.; Tanaka, S.; Isoda, F.; Kleopoulos, S.; Rosenfeld, M.; Kaplitt, M.; Sekihara, H.; Mobbs, C. |
| Article Title: | Defective adenoassociated viral-mediated transfection of insulin gene by direct injection into liver parenchyma decreases blood glucose of diabetic mice |
| Abstract: | The present study assessed the feasibility of transferring the insulin gene into liver cells of diabetic individuals using a defective adenoassociated vital (AAV) vehicle. AAV offers several advantages over other vital vectors, since this vehicle can facilitate transfection in vivo without cell division and without any viral coding sequences (thus minimizing inflammation). The rat insulin gene and lacZ were each packed into a defective AAV vehicle (AAV-INS and AAV-lacZ, respectively). Successful AAV- mediated transfection and expression of lacZ into hepatocytes in primary cell culture were demonstrated by chemiluminescent assay of beta-galactosidase. Similarly, AAV-mediated transfection and expression of the insulin gene into hepatocytes was demonstrated by immunocytochemistry and reverse-transcriptase polymerase chain reaction (RT-PCR). After AAV-mediated transfection of the insulin gene into hepatocytes, glucose in the medium was significantly reduced for up to 5 days. After direct injection of AAV-INS into liver parenchyma of diabetic mice, successful transfection was demonstrated by RT- PCR, and blood glucose was significantly decreased for at least 6 days. These studies suggest that the AAV vector may be used to transfer the insulin gene into liver cells in vitro and in vivo. |
| Keywords: | immunohistochemistry; controlled study; nonhuman; polymerase chain reaction; animal cell; mouse; animals; mice; cells, cultured; reverse transcription polymerase chain reaction; gene expression; cell line; animal experiment; animal model; transfection; gene transfer; animalia; genetic vectors; molecular cloning; liver; genetic transfection; immunocytochemistry; feasibility studies; diabetes mellitus; rat; gene therapy; insulin; glucose blood level; glucose; blood glucose; rats; rats, sprague-dawley; liver parenchyma; expression vector; beta-galactosidase; defective viruses; liver cell culture; diabetes mellitus, experimental; chemiluminescent measurements; adeno associated virus; adeno-associated virus; dependovirus; aav; male; priority journal; article; defective vector; streptozocin diabetes |
| Journal Title: | Hormone and Metabolic Research |
| Volume: | 29 |
| Issue: | 12 |
| ISSN: | 0018-5043 |
| Publisher: | Georg Thieme Verlag Kg |
| Date Published: | 1997-12-01 |
| Start Page: | 599 |
| End Page: | 603 |
| Language: | English |
| PUBMED: | 9497894 |
| PROVIDER: | scopus |
| DOI: | 10.1055/s-2007-979108 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 17 March 2017 -- Source: Scopus |
