tal protein was isolated and western blot was done to analysis the HCV core protein expression. Genomic DNA was isolated, and genomic integration was confirmed by nested PCR. The resultant bands were sequenced and aligned with the genomic sites. The switch from attB to genomic sequence near the TTG core and the detectable sequence identity between the genomic sequence and attP confirmed FC31- mediated integration at genomic pseudo-attP sites. These results further demonstrated that plasmid integration was associated with higher sustained levels of transgene expression. To investigate the shRNA hepatotoxicity, the mice were Tasquinimod citations injected with pSilencer-2.1-U6 plasmid, control non-targeting shRNA expression vectors, or shRNA523 expression vectors. Serum levels of alanine aminotransferase, a marker of liver function, were evaluated. ALT levels were significantly increased 8 h after injection, subsided to 167�C214 IU/L by 48 h, then declined to the baseline by 120 h. There were no significant difference observed Ganetespib citations across all groups. In agreement with the ALT observations, cytokine IL-6 levels in serum, which is essential for an optimal acute-phase response after tissue damage, were very high across each group 8 h post injection, subsiding to 26.00�C46.87 pg/ml by 48 h, with no significant difference observed for shRNA-Scramble, shRNA523 vs. vehicle treatment. Another proinflammatory cytokine IL-1b levels exhibited a rise 8 h after injection, followed by a return to the baseline levels during the next 48 hours. There was also no statistical significance between the groups. Examination of liver histology from both treated mice revealed significant hydrodynamic injection-related hepatic injury. At 8 h after injection liver morphology underwent remarkable changes. Many hepatocytes were swollen and their cytoplasm was vacuolized and stained less with eosin. Red blood cells appeared as clusters between and inside damaged hepatocytes. Cells developed signs of irreversible damage such as apoptosis or necrosis, accompanied by minimal neutrophil infiltration. Liver morphology 24 h after HTV injection was close to normal. Single cell necrosis, swollen cells and inflammatory infiltration were infrequent at 24 h, showing liver recovery at this time point. At 48 h the liver morphology became more normal. Taken together, these results indicated that liver damage observed in the mice was due to hydrodynamic injection, and all the mice could recover from hydrodynamic injection up to 2 days. We described here a novel method to screen anti-core protein siRNA in the mouse liver. By using the reporter gene, anti-core protein compounds can be screened by simply bioluminescence imaging the Fluc activity in whole animals under true physiological conditions. In this study, three shRNAs
