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
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).