New Bapx1(Cre-EGFP) mouse lines for lineage tracing and conditional knockout studies
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Published on: December 18, 2015
V Sivakamasundari1, Hsiao Yun Chan, Sook Peng Yap
1Stem Cell and Developmental Biology, Genome Institute of Singapore, Singapore 138672.
Abstract:
To gain insight into the roles of various genes in development and to circumvent embryonic lethality that hinders genetic studies, lineage tracing and conditional knockout techniques have been widely performed on mice using the increasing numbers of gene-targeted Cre mouse lines. Employing the internal ribosome entry site (IRES) and the 2A peptide multicistronic expression strategies, we report two new Bapx1 mouse lines with functional Bapx1 whereby Cre and enhanced green fluorescence protein (EGFP) are expressed discretely under the control of the Bapx1 promoter. These mouse lines, when mated with the Rosa26R-lacZ reporter line, can be used to trace the lineage of Bapx1-expressing cells whereas stage-specific, spatial expression of Bapx1 can be visualized by the EGFP fluorescence. In addition, both of our Bapx1(Cre-EGFP) mouse lines can be used to enrich for Bapx1-specific cells and also serve as effective conditional knockout tools to investigate gene functions in the skeleton and/or visceral organs.
Insights
We developed two new Bapx1 mouse lines for studying gene function. These lines enable lineage tracing and conditional knockout of Bapx1 (Bapx1-Cre-EGFP) to understand its role in development.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- Genetic studies are crucial for understanding gene function in development.
- Embryonic lethality often complicates genetic research in mice.
- Cre-lox technology and lineage tracing are vital tools in developmental studies.
Purpose of the Study:
- To create novel mouse models for studying the Bapx1 gene.
- To enable lineage tracing of Bapx1-expressing cells.
- To facilitate conditional knockout of Bapx1 for functional analysis.
Main Methods:
- Utilized internal ribosome entry site (IRES) and 2A peptide multicistronic expression strategies.
- Generated two new Bapx1(Cre-EGFP) mouse lines.
- Crossed Bapx1(Cre-EGFP) mice with Rosa26R-lacZ reporter mice.
Main Results:
- Developed Bapx1 mouse lines expressing Cre and enhanced green fluorescence protein (EGFP) under the Bapx1 promoter.
- Demonstrated utility for lineage tracing of Bapx1-expressing cells using the reporter line.
- Showcased EGFP fluorescence for visualizing stage-specific and spatial Bapx1 expression.
Conclusions:
- The new Bapx1(Cre-EGFP) mouse lines are effective tools for lineage tracing.
- These lines facilitate conditional knockout of Bapx1 to investigate gene function.
- The models aid in studying Bapx1's role in skeletal and visceral organ development.
