A collection of genetic mouse lines and related tools for inducible and reversible intersectional mis-expression
A collection of genetic mouse lines and related tools for inducible and reversible intersectional mis-expression
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DOI：
10.1242/dev.186650
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发表时间：
2020-05-01
期刊：
影响因子：
4.6
通讯作者：
Rosello-Diez, Alberto
中科院分区：
文献类型：
--
作者：
Ahmadzadeh, Elham;Bayin, N. Sumru;Rosello-Diez, Alberto
由于基因操作方面的许多进展，小鼠模型在询问生物过程的能力方面变得非常强大。为了精确定位特定细胞类型感兴趣的基因的表达，使用两个特定细胞类型特有的启动子/增强子的交叉遗传方法是理想的。在这些方法中，增加了对基因表达的时间控制的变体是最强大的。我们描述了一种涉及三个转基因的交叉方法的开发、验证和应用，该方法需要两个启动子/增强子的交叉来针对精确的细胞类型的基因表达。此外，该方法使用可用表达TTA/RTA的株系来控制基因表达的时间，分别基于是否存在多西胞苷。我们还表明，该方法可以扩展到其他动物模型，使用鸡胚胎。我们建立了三个以TIGRE(Igs7)为靶点、三个基因(p21、DTA和CTGF)上游带有tre-loxP-tdTomato-loxP的小鼠系，并将它们与靶向小脑和四肢表达的Cre和TTA/RTTA系相结合。我们的工具将有助于揭开多个领域和生物体中的生物学问题。
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 using 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 uses available lines expressing tTA/rTA to control the timing of gene expression based on whether doxycydine 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.