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A collection of genetic mouse lines and related tools for inducible and reversible intersectional misexpression

By Elham Ahmadzadeh, N. Sumru Bayin, Xinli Qu, Aditi Singh, Linda Madisen, Daniel Stephen, Hongkui Zeng, Alexandra L. Joyner, Alberto Rosello-Diez

Posted 05 Sep 2019
bioRxiv DOI: 10.1101/754192 (published DOI: 10.1242/dev.186650)

Thanks to many advances in genetic manipulation, mouse models have become very powerful in their ability to interrogate biological processes. In order to precisely target expression of a gene of interest to particular cell types, intersectional genetic approaches utilizing two promoter/enhancers unique to a cell type are ideal. Within these methodologies, variants that add temporal control of gene expression are the most powerful. We describe the development, validation and application of an intersectional approach that involves three transgenes, requiring the intersection of two promoter/enhancers to target gene expression to precise cell types. Furthermore, the approach utilizes available lines expressing tTA/rTA to control timing of gene expression based on whether doxycycline is absent or present, respectively. We also show that the approach can be extended to other animal models, using chicken embryos. We generated three mouse lines targeted at the Tigre ( Igs7 ) locus with TRE-loxP-tdTomato-loxP upstream of three genes ( p21, DTA and Ctgf ) and combined them with Cre and tTA/rtTA lines that target expression to the cerebellum and limbs. Our tools will facilitate unraveling biological questions in multiple fields and organisms. Summary statement Ahmadzadeh et al. present a collection of four mouse lines and genetic tools for misexpression-mediated manipulation of cellular activity with high spatiotemporal control, in a reversible manner.

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