Epigenetics & Neurobiology Unit, European Molecular Biology Laboratory (EMBL), Rome, Italy.
Systems Epigenetics, Otto Warburg Laboratories, Max Planck Institute for Molecular Genetics, Berlin, Germany.
Methods Mol Biol. 2024;2842:57-77. doi: 10.1007/978-1-0716-4051-7_3.
Epigenome editing has emerged as a powerful technique for targeted manipulation of the chromatin and transcriptional landscape, employing designer DNA binding domains fused with effector domains, known as epi-editors. However, the constitutive expression of dCas9-based epi-editors presents challenges, including off-target activity and lack of temporal resolution. Recent advancements of dCas9-based epi-editors have addressed these limitations by introducing innovative switch systems that enable temporal control of their activity. These systems allow precise modulation of gene expression over time and offer a means to deactivate epi-editors, thereby reducing off-target effects associated with prolonged expression. The development of novel dCas9 effectors regulated by exogenous chemical signals has revolutionized temporal control in epigenome editing, significantly expanding the researcher's toolbox. Here, we provide a comprehensive review of the current state of these cutting-edge systems and specifically discuss their advantages and limitations, offering context to better understand their capabilities.
表观基因组编辑技术已经成为一种强大的技术，可用于靶向修饰染色质和转录景观，采用与效应结构域融合的设计 DNA 结合结构域，称为表观遗传编辑工具。然而，基于 dCas9 的表观遗传编辑工具的组成型表达存在一些挑战，包括脱靶活性和缺乏时间分辨率。最近，基于 dCas9 的表观遗传编辑工具的进展通过引入创新的开关系统解决了这些限制，这些系统可以实现其活性的时间控制。这些系统允许在时间上精确调节基因表达，并提供了一种使表观遗传编辑工具失活的方法，从而减少与长时间表达相关的脱靶效应。受外源化学信号调节的新型 dCas9 效应子的开发革新了表观基因组编辑中的时间控制，极大地扩展了研究人员的工具包。在这里，我们全面回顾了这些前沿系统的现状，并特别讨论了它们的优缺点，为更好地理解它们的能力提供了背景。
