Recombinatorial Cloning to Generate Ready-for-injection Clones
2:23
Hydrodynamic Tail Vein Injection
4:22
Induction of Transfected CreER with Tamoxifen
5:33
Induction of Tetracycline-dependent Gene or shRNA Expression
6:35
Preparation of Mouse Liver for Analysis by Immunostaining
7:30
Results: Constitutive and Inducible Gene Expression in the Liver Following HTVI
8:47
Conclusion
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The overall goal of this method is the stable transfection of hepatocytes in vivo, with vector constructs for inducible gene or shRNA expression. This method can help to answer key questions in the field of liver research. For example, it can be a
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Hydrodynamic tail vein injection of transposon-based integration vectors enables stable transfection of murine hepatocytes in vivo. Here, we present a practical protocol for transfection systems that enables the long-term constitutive expression of a single transgene or combined constitutive and doxycycline-inducible expression of a transgene or miR-shRNA in the liver.