Generowanie myszy z nokautem specyficznych dla brązowego tłuszczu przy użyciu połączonego systemu Cre-LoxP, CRISPR-Cas9 i jednoprzewodnikowego systemu RNA wirusa związanego z adenowirusem

Overview

This protocol describes a method to generate brown adipose tissue (BAT)-specific knockout mice using a combination of Cre-LoxP, CRISPR-Cas9, and adeno-associated virus (AAV) single-guide RNA techniques. By utilizing these methods, researchers can quickly create tissue-specific knockouts without the need for selective breeding.

Key Study Components

Research Area

• Genetics
• Cell biology
• Molecular biology

Background

• Brown adipose tissue is crucial for energy metabolism.
• Creating tissue-specific knockouts aids in understanding biological processes.
• This method can potentially be adapted for other tissues.

Methods Used

• Application of Cre-LoxP and CRISPR-Cas9 approaches.
• Mouse model utilized: UCP1 Cre Cas9 mice.
• Administration of AAV-sgRNA via surgical microinjection.

Main Results

• Successful targeting and knockout of brown adipose tissue.
• Demonstrated effectiveness of AAV in tissue-specific gene editing.
• Key surgical techniques validated for precision delivery.

Conclusions

• The study showcases a rapid technique for generating knockout models pertinent to brown fat metabolism.
• It enhances understanding of molecular regulators in energy metabolism.

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