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Mapping Transgene Insertion Sites Reveals Complex Interactions Between Mouse Transgenes and Neighboring Endogenous
Mallory A Laboulaye1, Xin Duan1, Mu Qiao1
1Center for Brain Science and Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, United States.
Frontiers in Molecular Neuroscience
|November 9, 2018
Summary
Mapping transgene insertion sites using Targeted Locus Amplification (TLA) reveals that transgene expression can be influenced by nearby genes. This study shows transgenes impact neighboring gene expression, highlighting the importance of insertion site analysis for transgenic mouse models.
Area of Science:
- Genetics
- Molecular Biology
- Neuroscience
Background:
- Transgenic mouse lines are crucial for cell labeling and manipulation.
- Off-target gene expression in transgenic models is often linked to transgene integration sites.
- Understanding transgene-host gene interactions is vital for accurate experimental interpretation.
Purpose of the Study:
- To investigate the hypothesis that transgene insertion site influences expression patterns.
- To map the integration sites of three specific transgenic mouse lines (HB9-GFP, Mito-P, TYW3) in the retina.
- To determine the relationship between transgene expression and nearby endogenous genes.
Main Methods:
- Utilized Targeted Locus Amplification (TLA), a proximity ligation-based method, to identify transgene insertion sites.
- Analyzed the genomic proximity of transgenes to endogenous genes.
- Assessed gene expression patterns in relation to transgene location and neighboring genes.
Main Results:
- Identified nearest endogenous genes to HB9-GFP, Mito-P, and TYW3 transgenes as Cdh6, Fat4, and Khdrbs2, respectively.
- Demonstrated that transgene expression in two lines (Mito-P and HB9-GFP) correlated with the expression of their closest endogenous genes (Fat4 and Cdh6), even at distances of 550 and 680 kb.
- Observed that all three transgenes reduced the expression of their neighboring endogenous genes, which were expressed in cell types targeted by the respective transgenic lines.
Conclusions:
- Transgene insertion site mapping is essential for interpreting expression data from transgenic mouse lines.
- Transgenes can influence the expression of nearby endogenous genes, and vice versa.
- Targeted Locus Amplification (TLA) is a reliable technique for discovering transgene integration sites.
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