Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China.
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China; International Research Center for Marine Biosciences, Ministry of Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Gene. 2018 Nov 30;677:273-279. doi: 10.1016/j.gene.2018.07.077. Epub 2018 Aug 1.
CRISPR activation (CRISPRa) system is the convenient tool for targeted-gene activation, it has been developed and combined with a lighting-based system that can control transcription initiation spatially and temporally by utilizing photoreceptor derived from plant Arabidopsis thaliana. A blue light photoreceptor the Cryptochrome 2 (CRY2), and its binding partner CIB1 will dimerize by exposure to the blue light and it has been applied to human cells. However, the application of a combination of these two systems to zebrafish cell is still not explored. We performed zebrafish gene activation using p65 and VP64 activators in the zebrafish cells (ZF4). Our study demonstrated that we have successfully controlled the transcription level of ASCL1a, BCL6a, and HSP70 genes using blue light-activated CRISPR activation system. The result showed that using this system, mRNA level expression of ASCL1a, BCL6a, and HSP70 genes increased after irradiated under blue light for several hours and significantly different to those which treated in the dark.
CRISPR 激活（CRISPRa）系统是一种靶向基因激活的便捷工具，它已经被开发出来并与基于光照的系统相结合，该系统可以利用源自拟南芥的光受体来时空控制转录起始。蓝光光受体隐花色素 2（CRY2）及其结合伴侣 CIB1 在暴露于蓝光时会二聚化，并已应用于人类细胞。然而，这两个系统的组合在斑马鱼细胞中的应用尚未得到探索。我们在 ZF4 斑马鱼细胞中使用 p65 和 VP64 激活子进行了斑马鱼基因激活。我们的研究表明，我们已经成功地使用蓝光激活的 CRISPR 激活系统控制了 ASCL1a、BCL6a 和 HSP70 基因的转录水平。结果表明，使用该系统，在蓝光照射数小时后，ASCL1a、BCL6a 和 HSP70 基因的 mRNA 水平表达增加，与在黑暗中处理的基因相比有显著差异。
