Dubois Annick, Roudier François
Laboratoire Reproduction et Développement des Plantes, Université de Lyon, ENS de Lyon, UCB Lyon 1, CNRS, INRAE, 69364 Lyon, France.
Epigenomes. 2021 Aug 24;5(3):17. doi: 10.3390/epigenomes5030017.
CRISPR-based epigenome editing uses dCas9 as a platform to recruit transcription or chromatin regulators at chosen loci. Despite recent and ongoing advances, the full potential of these approaches to studying chromatin functions in vivo remains challenging to exploit. In this review we discuss how recent progress in plants and animals provides new routes to investigate the function of chromatin regulators and address the complexity of associated regulations that are often interconnected. While efficient transcriptional engineering methodologies have been developed and can be used as tools to alter the chromatin state of a locus, examples of direct manipulation of chromatin regulators remain scarce in plants. These reports also reveal pitfalls and limitations of epigenome engineering approaches that are nevertheless informative as they are often associated with locus- and context-dependent features, which include DNA accessibility, initial chromatin and transcriptional state or cellular dynamics. Strategies implemented in different organisms to overcome and even take advantage of these limitations are highlighted, which will further improve our ability to establish the causality and hierarchy of chromatin dynamics on genome regulation.
基于CRISPR的表观基因组编辑利用dCas9作为平台，在选定的位点招募转录或染色质调节因子。尽管最近取得了进展且相关研究仍在进行，但在体内利用这些方法全面研究染色质功能仍具有挑战性。在本综述中，我们讨论了植物和动物领域的最新进展如何为研究染色质调节因子的功能提供新途径，并解决通常相互关联的相关调控的复杂性。虽然已经开发出了高效的转录工程方法，可作为改变基因座染色质状态的工具，但在植物中直接操纵染色质调节因子的例子仍然很少。这些报告还揭示了表观基因组工程方法的缺陷和局限性，不过它们仍具有参考价值，因为它们通常与位点和背景依赖的特征相关，这些特征包括DNA可及性、初始染色质和转录状态或细胞动力学。文中强调了在不同生物体中为克服甚至利用这些局限性而实施的策略，这将进一步提高我们确定染色质动力学对基因组调控的因果关系和层级关系的能力。
