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

Published: April 30, 2023

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.   Prepare sterile 0.9% saline for injection (do not use PBS). Use volu…

Açıklamalar

The authors have nothing to disclose.

Materials

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

Etiketler

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Bu Makaleden Alıntı Yapın
Hydrodynamic Gene Delivery: A Technique for In Vivo Transfection of Exogenous DNA in Murine Hepatocytes via Tail Vein Injection. J. Vis. Exp. (Pending Publication), e20990, doi: (2023).

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