Conditional gene modification in mouse liver using hydrodynamic delivery of plasmid DNA encoding Cre recombinase |
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Authors: | Zhu Huan Zhang Wang Wei Feng Deng Min Sai Yin Xue Jing Lun |
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Institution: | State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China. HZ_Zhu@hotmail.com |
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Abstract: | The success of Cre-mediated conditional gene targeting in liver of mice has until now depended on the generation of Cre recombinase transgenic mice or on viral-mediated transduction. Here, we sought to establish the feasibility of using hydrodynamic gene delivery of Cre recombinase into liver, using a ROSA26 EGFP mouse. The expression of EGFP and beta-galactosidase was exclusively detected in the liver of mice treated with hydrodynamic gene delivery of Cre recombinase, as assessed with fluorescence microscopy and X-Gal staining, respectively; Southern blotting also showed that Cre mediated recombination occurred specifically in the liver and not in other organs. The Cre mediated recombination reached about 61% of hepatocytes of mouse after repeated injection, as analyzed by flow cytometry. These results demonstrate that Cre recombinase can be transferred to the liver of mice through a simple hydrodynamic gene-delivery approach and can mediate efficient recombination in hepatocytes. Thus, hydrodynamic gene delivery of the Cre recombinase provides a valuable approach for Cre-loxP-mediated conditional gene modification in the liver of mice. |
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Keywords: | Alb e/p albumin enhancer and promoter ALT alanine aminotransferase AST aspartate aminotransferase CMV cytomegalovirus EGFP enhancer green fluorescent protein 4-OHT 4-hydroxytamoxifen PBS phosphate-buffered saline |
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