The integration site of the APP transgene in the J20 mouse model of Alzheimer's disease.
Justin L ToshMatthew RickmanEllie RhymesFrances E NoronaEmma ClaytonLennart MuckeAdrian M IsaacsElizabeth M C FisherFrances K WisemanPublished in: Wellcome open research (2017)
Background: Transgenic animal models are a widely used and powerful tool to investigate human disease and develop therapeutic interventions. Making a transgenic mouse involves random integration of exogenous DNA into the host genome that can have the effect of disrupting endogenous gene expression. The J20 mouse model of Alzheimer's disease (AD) is a transgenic overexpresser of human APP with familial AD mutations and has been extensively utilised in preclinical studies and our aim was to determine the genomic location of the J20 transgene insertion. Methods: We used a combination of breeding strategy and Targeted Locus Amplification with deep sequencing to identify the insertion site of the J20 transgene array. To assess RNA and protein expression of Zbtb20, we used qRT-PCR and Western Blotting. Results: We demonstrate that the J20 transgene construct has inserted within the genetic locus of endogenous mouse gene Zbtb20 on chromosome 16 in an array , disrupting expression of mRNA from this gene in adult hippocampal tissue, while expression of Zbtb20 protein remains unchanged. We note that the endogenous mouse App gene also lies on chromosome 16, although 42 Mb from the Zbtb20 locus. Conclusions: These data will be useful for future studies utilising this popular model of AD, particularly those investigating gene interactions between the J20 APP transgene and other genes present on Mmu16 in the mouse.
Keyphrases
- copy number
- genome wide
- mouse model
- genome wide identification
- gene expression
- dna methylation
- endothelial cells
- poor prognosis
- binding protein
- genome wide analysis
- high resolution
- cognitive decline
- high throughput
- physical activity
- transcription factor
- small molecule
- big data
- mass spectrometry
- genome wide association study
- circulating tumor
- machine learning
- induced pluripotent stem cells
- cell free
- cancer therapy
- mesenchymal stem cells
- single molecule
- bone marrow
- current status
- artificial intelligence
- real time pcr