Hydrodynamic Gene Delivery: A Technique for In Vivo Transfection of Exogenous DNA in Murine Hepatocytes via Tail Vein Injection

Abstract

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).

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.

Protocol

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.  

1. Prepare sterile 0.9% saline for injection (do not use PBS). Use volume corresponding to about 10% of mouse body weight. Example: for a mouse weighing 20 g, prepare 2 mL of solution.
2. Prepare injection vectors that we....

Materials

List of materials used in this article

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