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Biology
Hydrodynamische Nierenbecken Injektion für nicht-virale Expression von Proteinen in der Niere
Hydrodynamische Nierenbecken Injektion für nicht-virale Expression von Proteinen in der Niere
JoVE Journal
Biology
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JoVE Journal Biology
Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney

Hydrodynamische Nierenbecken Injektion für nicht-virale Expression von Proteinen in der Niere

Full Text
19,713 Views
08:26 min
January 8, 2018

DOI: 10.3791/56324-v

Lauren E. Woodard1,2,3, Richard C. Welch2, Felisha M. Williams2, Wentian Luo2, Jizhong Cheng3, Matthew H. Wilson1,2,3

1Department of Veterans Affairs,Tennessee Valley Healthcare System, 2Departments of Medicine and Pharmacology,Vanderbilt University Medical Center, 3Department of Medicine,Baylor University College of Medicine

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol describes a method to inject plasmid DNA into the mouse kidney via the renal pelvis to produce transgene expression specifically in the kidney. This technique allows for reproducible protein expression in the adult mouse kidney, addressing key questions in nephrology.

Key Study Components

Area of Science

  • Nephrology
  • Gene Therapy
  • Molecular Biology

Background

  • Understanding protein expression in the kidney is crucial for nephrology research.
  • Existing methods may lead to gene expression in multiple organs.
  • This technique focuses on specific expression in the kidney.
  • Visual demonstration is essential for successful execution of the method.

Purpose of Study

  • To produce protein expression from plasmid DNA in the adult mouse kidney.
  • To investigate the effects of specific proteins on kidney function.
  • To provide a reliable method for nephrology research.

Methods Used

  • Resuspension of plasmid DNA in hydrodynamic delivery buffer.
  • Placement of the mouse on a water circulated heat pad.
  • Injection of plasmid DNA into the renal pelvis.
  • Careful exposure and quick execution of the injection.

Main Results

  • Successful transgene expression in the kidney.
  • Reproducibility of protein expression demonstrated.
  • Specific targeting of the kidney achieved.
  • Method validated for further nephrology studies.

Conclusions

  • This method provides a new tool for nephrology research.
  • It allows for targeted gene expression in the kidney.
  • Visual guidance enhances the success of the procedure.

Frequently Asked Questions

What is the main advantage of this injection method?
The main advantage is that it produces protein expression specifically in the kidney, avoiding expression in other organs.
Why is visual demonstration important?
Visual demonstration is critical as the injection requires careful exposure and must be done quickly for effective DNA entry into cells.
What is the purpose of resuspending plasmid DNA?
Resuspending plasmid DNA in hydrodynamic delivery buffer prepares it for injection into the renal pelvis.
How does this method contribute to nephrology research?
It allows researchers to study the effects of specific proteins on kidney function in a controlled manner.
What type of animal is used in this protocol?
The protocol uses adult mice for the injection of plasmid DNA.
Is this method reproducible?
Yes, the method has been shown to produce reproducible results in protein expression.

Dieses Protokoll beschreibt eine Methode zum Injizieren von Plasmid DNA in der Mausniere über das Nierenbecken, Transgene Ausdruck speziell in der Niere zu produzieren.

Das übergeordnete Ziel dieses chirurgischen Eingriffs ist es, die Proteinexpression aus Plasmid-DNA in der adulten Mausniere zu erzeugen. Diese Methode ist ein neues Werkzeug, um Schlüsselfragen in der Nephrologie zu beantworten, z. B. wie sich die Expression eines interessierenden Proteins auf die adulte Niere auswirkt. Der Hauptvorteil dieser Technik besteht darin, dass sie die Proteinexpression auf reproduzierbare Weise erzeugt, insbesondere in der Niere, im Gegensatz zu anderen Methoden, die auch in anderen Organen eine Genexpression erzeugen können.

Die visuelle Demonstration dieser Methode ist von entscheidender Bedeutung, da die Injektion des Nierenbeckens eine sorgfältige Exposition erfordert und schnell erfolgen muss, damit die Plasma-DNA in die Zellen gelangen kann. Beginnen Sie mit der Resuspendierung der Plasmid-DNA im hydrodynamischen Verabreichungspuffer. Legen Sie die Maus auf ein wasserumgewälztes Heizkissen, das mit einem blauen Pad bedeckt ist.

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