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Updated: Jun 18, 2025
12:04
Mouse Genome Engineering Using Designer Nucleases
Published on: April 2, 2014
28.8K
Mutagenesis on a complex mouse genetic background by site-specific nucleases.
Benjamin Davies1,2, Lucy Trelfa3,1, Victoria S Rashbrook3,1
1Wellcome Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, UK.
Transgenic Research
|August 1, 2024
Summary
Site-specific nucleases efficiently create genetic mutations in complex mouse models for atherosclerosis research. This method bypasses lengthy backcrossing, accelerating gene function studies and offering animal welfare benefits.
Area of Science:
- Genetics
- Cardiovascular Research
- Preclinical Models
Background:
- Complex mouse models are crucial for preclinical research but require time-consuming backcrossing.
- Developing new genetic modifications on existing complex backgrounds is challenging.
Purpose of the Study:
- To evaluate the efficacy of site-specific nucleases for generating mutations in a complex genetic background.
- To investigate the role of ATP binding cassette transporter G1 (ABCG1) in atherosclerosis regression using this method.
Main Methods:
- Utilized the REVERSA mouse model for atherosclerosis, which has four genetic alterations.
- Microinjected site-specific nucleases into zygotes from the REVERSA model to create ABCG1 knockouts.
- Assessed cholesterol efflux in bone marrow-derived macrophages and atherosclerosis regression in the aortic root and arch.
Main Results:
- Successfully generated two independent ABCG1 knockout lines on the REVERSA background.
- Observed a significant reduction in cholesterol efflux to HDL in macrophages from knockout mice.
- Found no impact of Abcg1 loss on atherosclerosis regression in the aortic root or arch.
Conclusions:
- Site-specific nucleases are effective for creating genetic modifications directly on complex disease backgrounds.
- This approach facilitates gene function exploration without extensive backcrossing, aligning with the 3Rs principles.
- ABCG1 does not play a significant role in atherosclerosis regression in this model.
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