Bao Zehua, Jain Surbhi, Jaroenpuntaruk Valerie, Zhao Huimin
Department of Biochemistry, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
Departments of Chemical and Biomolecular Engineering, Chemistry, and Bioengineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
ACS Synth Biol. 2017 Apr 21;6(4):686-693. doi: 10.1021/acssynbio.6b00313. Epub 2017 Jan 19.
The concerted action of multiple genes in a time-dependent manner controls complex cellular phenotypes, yet the temporal regulation of gene expressions is restricted on a single-gene level, which limits our ability to control higher-order gene networks and understand the consequences of multiplex genetic perturbations. Here we developed a system for temporal regulation of multiple genes. This system combines the simplicity of CRISPR/Cas9 activators for orthogonal targeting of multiple genes and the orthogonality of chemically induced dimerizing (CID) proteins for temporal control of CRISPR/Cas9 activator function. In human cells, these transcription activators exerted simultaneous activation of multiple genes and orthogonal regulation of different genes in a ligand-dependent manner with minimal background. We envision that our system will enable the perturbation of higher-order gene networks with high temporal resolution and accelerate our understanding of gene-gene interactions in a complex biological setting.
多个基因以时间依赖性方式协同作用控制复杂的细胞表型，然而基因表达的时间调控在单基因水平上受到限制，这限制了我们控制高阶基因网络以及理解多重基因扰动后果的能力。在此，我们开发了一种用于多个基因时间调控的系统。该系统结合了CRISPR/Cas9激活剂对多个基因进行正交靶向的简便性以及化学诱导二聚化（CID）蛋白对CRISPR/Cas9激活剂功能进行时间控制的正交性。在人类细胞中，这些转录激活剂以配体依赖性方式同时激活多个基因，并以最小的背景对不同基因进行正交调控。我们设想，我们的系统将能够以高时间分辨率扰动高阶基因网络，并加速我们对复杂生物学环境中基因-基因相互作用的理解。
