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Biology
肾水动力肾盂注射非病毒蛋白在肾脏的表达
肾水动力肾盂注射非病毒蛋白在肾脏的表达
JoVE Journal
Biology
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JoVE Journal Biology
Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney

肾水动力肾盂注射非病毒蛋白在肾脏的表达

Full Text
19,707 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.

本协议描述了通过肾盂向小鼠肾脏注入质粒 DNA 的方法, 以在肾脏中特异性地产生转基因表达。

该外科手术的总体目标是从成年小鼠肾脏中的质粒 DNA 中产生蛋白质表达。这种方法是解决肾脏病学中关键问题的新工具,例如,目标蛋白质的表达如何影响成人肾脏。该技术的主要优点是它以可重复的方式产生蛋白质表达,特别是在肾脏中,这与其他方法可能在其他器官中产生基因表达不同。

这种方法的视觉演示至关重要,因为肾盂注射需要仔细暴露,并且必须快速完成才能使血浆 DNA 进入细胞。首先在流体动力学递送缓冲液中重悬质粒 DNA。将鼠标放在覆盖有蓝色垫子的水循环加热垫上。

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