Abstract
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. Preliminary data suggests that ZBTB20 protein levels remain unchanged in this tissue, however further study is necessary. 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.
Highlights
The Tg(PDGFB-APPSwInd)20Lms (MGI:3057148, here referred to as ‘J20’) mouse model is a transgenic animal that overexpresses mutant human amyloid beta protein precursor (APP) protein, and is widely used as a model of amyloid deposition and pathogenesis in the study of Alzheimer’s disease (AD)
Mouse breeding strategy To investigate the role of Chmp2b in the pathogenesis of AD, we attempted to generate a homozygous knockout of Chmp2b in the Tg(PDGFB-APPSwInd)20Lms (J20) APP transgenic mouse strain
To accomplish this we set up matings over two generations (Figure 1a), firstly between Chmp2b-/- and hemizygous J20 mice
Summary
The Tg(PDGFB-APPSwInd)20Lms (MGI:3057148, here referred to as ‘J20’) mouse model is a transgenic animal that overexpresses mutant human APP protein (amyloid precursor protein), and is widely used as a model of amyloid deposition and pathogenesis in the study of Alzheimer’s disease (AD). The model was developed by Mucke and colleagues using the PDGF-APPSw,Ind transgene construct described previously (Games et al, 1995; Rockenstein et al, 1995), which includes a human APP mini-gene, carrying the familial AD-linked 717V-F (Indiana) mutation (Murrell et al., 1991) and. The transgene construct was designed so that the APP mini-gene included genomic sequence for APP introns 6–8, allowing expression of hAPP695, hAPP751 and hAPP770 isoforms. APPSw,Ind transgene expression is driven in neurons throughout the brain by the human platelet-derived growth factor β chain (PDGFβ) promoter (Harris et al, 2010; Sasahara et al, 1991). The J20 mouse is an important model: currently, 125 articles have been catalogued in the Mouse Genome
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