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Rapid Knockout and Reporter Mouse Line Generation and Breeding Colony Establishment Using EUCOMM Conditional-Ready
James L J Coleman1, Karen Brennan2, Tony Ngo1
1Molecular Cardiology Program, Victor Chang Cardiac Research Institute , Darlinghurst, NSW , Australia ; St. Vincent's Clinical School, University of New South Wales , Darlinghurst, NSW , Australia.
Frontiers in Endocrinology
|July 16, 2015
Summary
Researchers can now efficiently generate knockout mouse models for genetic studies. The International Knockout Mouse Consortium provides accessible embryonic stem cells, streamlining the creation of global and conditional knockout mice.
Area of Science:
- Genetics
- Genomics
- Mouse Models
Background:
- Generating knockout mouse lines was historically costly and time-intensive.
- The International Knockout Mouse Consortium (IKMC) established an open-access repository of embryonic stem (ES) cells in 2007.
- This repository facilitates gene deletion in mouse models through recombinase technology.
Purpose of the Study:
- To describe the experience of using the IKMC repository for creating mouse lines.
- To detail the process of obtaining germline transmission of European Conditional Mouse Mutagenesis Program (EUCOMM) constructs.
- To outline strategies for generating global and conditional knockouts and reporter mice.
Main Methods:
- Utilizing the IKMC's open-access ES cell repository.
- Employing sequential breeding with FLP1 and Cre recombinase transgenic mice.
- Implementing a specific breeding strategy for generating knockout and reporter mouse lines.
Main Results:
- Successfully obtained germline transmission of two EUCOMM "knockout-first" gene targeted constructs.
- Generated heterozygous knockout mice within six months of initiating breeding.
- Identified advantages and challenges associated with the EUCOMM system.
Conclusions:
- The IKMC repository significantly simplifies the generation of versatile knockout mouse lines.
- The described approach assists researchers in applying this valuable resource for experimental purposes.
- Efficient generation of knockout mouse models is now more accessible to the research community.
