Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, OX3 9DS, UK.
School of Biological Sciences, Universiti Sains Malaysia, Penang, 11800, Malaysia.
Nat Commun. 2017 Nov 21;8(1):1663. doi: 10.1038/s41467-017-01592-3.
Binary expression systems have revolutionised genetic research by enabling delivery of loss-of-function and gain-of-function transgenes with precise spatial-temporal resolution in vivo. However, at present, each existing platform relies on a defined exogenous transcription activator capable of binding a unique recognition sequence. Consequently, none of these technologies alone can be used to simultaneously target different tissues or cell types in the same organism. Here, we report a modular system based on programmable transcription activator-like effector (TALE) proteins, which enables parallel expression of multiple transgenes in spatially distinct tissues in vivo. Using endogenous enhancers coupled to TALE drivers, we demonstrate multiplexed orthogonal activation of several transgenes carrying cognate variable activating sequences (VAS) in distinct neighbouring cell types of the Drosophila central nervous system. Since the number of combinatorial TALE-VAS pairs is virtually unlimited, this platform provides an experimental framework for highly complex genetic manipulation studies in vivo.
二元表达系统通过在体内实现具有精确时空分辨率的基因敲除和功能获得转基因的传递，彻底改变了遗传研究。然而，目前，每种现有的平台都依赖于一种定义明确的外源性转录激活因子，该因子能够结合独特的识别序列。因此，这些技术都不能单独用于在同一生物体中同时靶向不同的组织或细胞类型。在这里，我们报告了一种基于可编程转录激活因子样效应蛋白（TALE）的模块化系统，该系统能够在体内空间上区分不同组织的多个转基因的平行表达。我们使用与 TALE 驱动子偶联的内源性增强子，在果蝇中枢神经系统的不同邻近细胞类型中演示了携带同源可变激活序列（VAS）的多个转基因的多路正交激活。由于组合 TALE-VAS 对的数量实际上是无限的，因此该平台为体内进行非常复杂的遗传操作研究提供了一个实验框架。
