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Generation of Brown Fat-Specific Knockout Mice Using a Combined Cre-LoxP, CRISPR-Cas9, and Adeno-Associated Virus Single-Guide RNA System
JoVE Journal
Biologia
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JoVE Journal Biologia
Generation of Brown Fat-Specific Knockout Mice Using a Combined Cre-LoxP, CRISPR-Cas9, and Adeno-Associated Virus Single-Guide RNA System

Generation of Brown Fat-Specific Knockout Mice Using a Combined Cre-LoxP, CRISPR-Cas9, and Adeno-Associated Virus Single-Guide RNA System

DOI:
10.3791/65083-v

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03:58 min

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March 24, 2023

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, ,
1Section on Integrative Physiology and Metabolism, Research Division, Joslin Diabetes Center,Harvard Medical School

Capitoli

  • 00:04Introduction
  • 00:55Surgery for Adeno‐Associated Virus (AAV) Injection into the Brown Adipose Tissue (BAT) in Mice
  • 02:41Results: Identifying the Potential Flaws Leading to Unsuccessful Injection
  • 03:18Conclusion

Summary

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In this protocol, we describe the technical procedures to generate brown adipose tissue (BAT)-specific knockout mice leveraging a combined Cre-LoxP, CRISPR-Cas9, and adeno-associated virus (AAV) single-guide RNA (sgRNA) system. The described steps include the design of the sgRNAs, the preparation of the AAV-sgRNA particles, and the microinjection of AAV into the BAT lobes.

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