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Protocol to knock in or out target genes in mouse epidydimal white adipose tissue by spot injecting adeno-associated

Wei Zhao1, Rexiati Ruze2, Xianghong Xie1

  • 1State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences & School of Basic Medicine Peking Union Medical College, Beijing 100005, China.

STAR Protocols
|November 17, 2022
PubMed

Insights

This study presents a method for gene editing in visceral fat tissue using adeno-associated viruses (AAV) in mice. This technique allows researchers to study the function of visceral fat in metabolic disease.

Area of Science:

  • Metabolic disease research
  • Gene editing techniques
  • Adipose tissue biology

Background:

  • Visceral fat obesity is linked to ectopic fat deposition, lipotoxicity, and metabolic disease.
  • Understanding visceral fat function is crucial for metabolic health.
  • Current methods for studying visceral fat gene function are limited.

Purpose of the Study:

  • To describe a protocol for gene knock-in or knock-out in mouse visceral fat tissue.
  • To provide a method for studying gene function in epididymal white adipose tissue (eWAT).
  • To enable efficient expression of exogenous proteins in mouse eWAT.

Main Methods:

  • Spot injection of adeno-associated viruses (AAV) into visceral fat tissue of live anesthetized mice.
  • Detailed procedures for anesthesia, incision, and injection into eWAT.
  • Techniques for efficient exogenous protein expression in mouse eWAT.

Main Results:

  • Successful gene manipulation (knock-in/knock-out) in visceral fat tissue.
  • Demonstration of efficient exogenous protein expression in mouse eWAT.
  • A reproducible protocol for studying visceral fat gene function.

Conclusions:

  • The described AAV spot injection technique is an effective method for genetic manipulation of visceral fat.
  • This protocol facilitates research into the role of visceral fat in metabolic disease.
  • The method allows for targeted gene function studies in eWAT.
Keywords:
MetabolismModel OrganismsMolecular Biology

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