Un abonnement à JoVE est nécessaire pour voir ce contenu. Connectez-vous ou commencez votre essai gratuit.
This video describes the technique of genetic in vivo transfection of murine hepatocytes by hydrodynamic tail vein injection. This transfection system generates a long-term expression of the transgene in the hepatocytes.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Preparing Solution for Hydrodynamic Tail Vein Injection
NOTE: Prepare the plasmid constructs for constitutive and inducible gene expression by cloning technique using competent bacteria.
2. Performing Hydrodynamic Tail Vein Injection
Name | Company | Catalog Number | Comments |
NaCl 0,9% | Braun | 3200905 | Carrier for intravenous injections |
Falcon Conical Tube 50ml | Corning Life Science | 352095 | |
Infrared Lamp | N/A | N/A | For warming of mouse tail |
Plasmids for cloning of sleeping beauty-transposon vectors for HTVI. | |||
pTC | n/a | Vector for constitutive gene expression | |
pEN_TTmcs | Addgene | #25755 | Entry vector for inducible gene expression |
pEN_TTGmiRc2 | Addgene | #25753 | Entry vector for inducible miRshRNA expression with coexpression of GFP |
pEN_TTmiRc2 | Addgene | #25752 | Entry vector for inducible miRshRNA expression without coexpression of GFP |
pTC ApoE-Tet | Addgene | #85578 | Expression vector for inducible gene or miR-shRNA expression with ApoE.HCR.hAAT promotor |
pTC-CMV-Tet | Addgene | #85577 | Expression vector for inducible gene or miR-shRNA expression with CMV promotor |
This article has been published
Video Coming Soon
Source: Hubner, E. K. et al., Constitutive and Inducible Systems for Genetic In Vivo Modification of Mouse Hepatocytes Using Hydrodynamic Tail Vein Injection. J. Vis. Exp. (2018).