Given the large amount of genome-wide data that has been collected during the last decades a good understanding of how and why cells change during development, homeostasis and disease might be expected. Unfortunately, the opposite is true; Triggers that cause cellular state changes remain elusive and the underlying molecular mechanisms are poorly understood. Although genes with the potential to influence cell states are known, the historic dependency on methods that manipulate gene expression outside the endogenous chromatin context has prevented us from understanding how cells organize, interpret and protect cellular programs. Fortunately, recent methodological innovations are now providing options to answer these outstanding questions, by allowing to target and manipulate individual genomic and epigenomic loci. In particular, three experimental approaches are now feasible due to DNA targeting tools: namely, activation and/or repression of master transcription factors in their endogenous chromatin context, targeting transcription factors to endogenous, alternative or inaccessible sites; and finally, functional manipulation of the chromatin context. In this article, we discuss the molecular basis of DNA targeting tools and review the potential of these new technologies before we summarize how these have already been used for the manipulation of cellular states and hypothesize about future applications.

中文翻译：

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用于生理学和细胞状态变化的CRISPR工具-转录工程和表观基因组编辑的潜力。

考虑到过去几十年来收集的大量全基因组数据，对细胞在发育，体内稳态和疾病过程中如何以及为何发生变化的很好理解。不幸的是，情况恰恰相反。引起细胞状态变化的触发因素仍然难以捉摸，其潜在的分子机制了解甚少。尽管已知具有潜在影响细胞状态的基因，但是历史上对内源性染色质环境之外操纵基因表达的方法的依赖使我们无法了解细胞如何组织，解释和保护细胞程序。幸运的是，最近的方法创新通过允许靶向和操纵单个基因组和表观基因组位点，提供了回答这些悬而未决问题的选择。尤其是，由于DNA靶向工具的存在，三种实验方法现在是可行的：即，在其内源染色质环境中激活和/或抑制主转录因子，将转录因子靶向内源性，替代性或不可及的位点；最后是染色质环境的功能操纵。在本文中，我们讨论了DNA靶向工具的分子基础，并回顾了这些新技术的潜力，然后总结了这些新技术如何已用于细胞状态的操纵以及未来应用的假设。染色质上下文的功能操纵。在本文中，我们讨论了DNA靶向工具的分子基础，并回顾了这些新技术的潜力，然后总结了这些新技术如何已用于细胞状态的操纵以及未来应用的假设。染色质上下文的功能操纵。在本文中，我们讨论了DNA靶向工具的分子基础，并回顾了这些新技术的潜力，然后总结了这些新技术如何已用于细胞状态的操纵以及未来应用的假设。