Konstitutive und induzierbaren Systeme für genetische In Vivo Änderung der Maus Hepatozyten mit hydrodynamischen Tail Vene Injektion

Overview

This article presents a method for stable transfection of murine hepatocytes in vivo using hydrodynamic tail vein injection of transposon-based integration vectors. The technique allows for long-term expression of transgenes or miR-shRNA, which can be induced by doxycycline.

Key Study Components

Area of Science

• Neuroscience
• Gene therapy
• Liver research

Background

• Stable transfection is crucial for studying gene function in vivo.
• Hepatocytes play a key role in liver function and disease.
• Inducible gene expression systems allow for temporal control of gene activity.
• Hydrodynamic injection is a technique used to deliver DNA to liver cells effectively.

Purpose of Study

• To establish a reliable method for stable transfection of hepatocytes.
• To enable the study of liver cancer mechanisms and regeneration.
• To facilitate the combined expression of CreER and inducible transgenes or shRNA.

Methods Used

• Hydrodynamic tail vein injection of transposon-based vectors.
• Combination of entry vector with short hairpin RNA and pTC Tet vector.
• Use of TE buffer for optimal vector preparation.
• Demonstration of the procedure by a laboratory med student.

Main Results

• Successful stable transfection of murine hepatocytes achieved.
• Inducible expression of transgenes and shRNA confirmed.
• Method demonstrated potential for liver research applications.
• Combined expression of CreER and transgenes was effective.

Conclusions

• The hydrodynamic tail vein injection method is effective for hepatocyte transfection.
• This technique can advance research in liver cancer and regeneration.
• Future studies can leverage this method for various gene expression investigations.

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