Division of Integrated Sciences for Life, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan.
Methods Mol Biol. 2023;2577:211-226. doi: 10.1007/978-1-0716-2724-2_15.
Genome editing technologies can be diverted into artificial transcription activators. In particular, researchers have improved dCas9-based technologies by tandem-fusing or trans-accumulating effector domains. Previously, we developed a hierarchical effector accumulation system named "TREE," enabling robust activation of target genes even when strongly silenced. In this chapter, we describe our protocol to design, construct, and validate the TREE-mediated target gene activation in cultured human cells.
基因组编辑技术可以转化为人工转录激活剂。特别是，研究人员通过串联融合或转位积累效应结构域改进了基于 dCas9 的技术。此前，我们开发了一种名为“TREE”的分层效应结构域积累系统，即使在强烈沉默的情况下，也能实现靶基因的稳健激活。在本章中，我们将描述设计、构建和验证 TREE 介导的培养人细胞中靶基因激活的方案。
