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Protocol for adeno-associated virus-mediated miRNA delivery in a rat heart failure model

Iqra Mushtaq1, Yu-Hsun Kao2, Yi-Jen Chen3

  • 1International Ph.D. Program in Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.

STAR Protocols
|December 13, 2024
PubMed

Insights

This study details a protocol for delivering microRNA (miRNA) to the heart using adeno-associated virus serotype 9 (AAV9) and hydrodynamic transfusion in rats with heart failure. The method enables effective miRNA delivery and expression analysis in cardiac tissue.

Area of Science:

  • Biomedical Engineering
  • Molecular Biology
  • Cardiovascular Research

Background:

  • Effective delivery of therapeutic microRNAs (miRNAs) to the heart is crucial for treating cardiac diseases.
  • Current methods face challenges in targeting the affected organ efficiently.
  • Adeno-associated virus (AAV) vectors show promise for gene therapy in the heart.

Purpose of the Study:

  • To present a detailed protocol for miRNA delivery to the failing heart in a rat model.
  • To establish a reliable method for assessing miRNA expression in cardiac tissue post-delivery.

Main Methods:

  • Induction of heart failure in a rat model.
  • Application of adeno-associated virus serotype 9 (AAV9) via hydrodynamic transfusion.
  • Echocardiography for cardiac function assessment.
  • Stem-loop RT-qPCR for miRNA expression analysis in cardiac tissue.

Main Results:

  • The protocol successfully delivers miRNA to the heart in a rat model of heart failure.
  • The method allows for quantitative assessment of miRNA expression in cardiac tissue.
  • Adeno-associated virus serotype 9 (AAV9) coupled with hydrodynamic transfusion is an effective delivery strategy.

Conclusions:

  • This protocol provides a robust method for cardiac miRNA delivery and evaluation.
  • The described technique can be valuable for preclinical studies of miRNA-based cardiac therapies.
  • Efficient miRNA delivery is key to developing novel treatments for heart failure.
Keywords:
cell biologygene expressionmodel organismsmolecular biologymolecular/chemical probes

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