Martella Andrea, Firth Mike, Taylor Benjamin J M, Göppert Anne, Cuomo Emanuela M, Roth Robert G, Dickson Alan J, Fisher David I
Discovery Biology, Discovery Sciences , R&D, AstraZeneca , Cambridge CB4 0WG , U.K.
Data Sciences and Quantitative Biology, Discovery Sciences , R&D, AstraZeneca , Cambridge CB4 0WG , U.K.
ACS Synth Biol. 2019 Sep 20;8(9):1998-2006. doi: 10.1021/acssynbio.8b00527. Epub 2019 Aug 21.
The ability to manipulate the expression of mammalian genes using synthetic transcription factors is highly desirable in both fields of basic research and industry for diverse applications, including stem cell reprogramming and differentiation, tissue engineering, and drug discovery. Orthogonal CRISPR systems can be used for simultaneous transcriptional upregulation of a subset of target genes while downregulating another subset, thus gaining control of gene regulatory networks, signaling pathways, and cellular processes whose activity depends on the expression of multiple genes. We have used a rapid and efficient modular cloning system to build and test in parallel diverse CRISPRa and CRISPRi systems and develop an efficient orthogonal gene regulation system for multiplexed and simultaneous up- and downregulation of endogenous target genes.
在基础研究和工业领域的各种应用中，包括干细胞重编程与分化、组织工程和药物发现等，利用合成转录因子来操控哺乳动物基因表达的能力是非常可取的。正交CRISPR系统可用于同时上调一部分靶基因的转录，而下调另一部分靶基因，从而实现对基因调控网络、信号通路以及其活性依赖于多个基因表达的细胞过程的控制。我们使用了一种快速高效的模块化克隆系统，并行构建和测试多种CRISPRa和CRISPRi系统，并开发了一种高效的正交基因调控系统，用于对内源靶基因进行多重同时上调和下调。
