CRISPR-mediated Synergistic Epigenetic and Transcriptional Control
CRISPR-mediated Synergistic Epigenetic and Transcriptional Control
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DOI：
10.1089/crispr.2021.0099
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发表时间：
2022-03-10
期刊：
影响因子：
3.7
通讯作者：
Qi, Lei S.
中科院分区：
文献类型：
--
作者：
Dominguez, Antonia A.;Chavez, Michael G.;Qi, Lei S.
靶向激活内源性基因是细胞工程的重要途径。在这里，我们报道了核酸酶失活的dCas9与转录激活因子(VPR)和表观遗传效应因子(组蛋白乙酰转移酶p300的催化结构域(Core))同时、顺序或作为单个四方效应因子融合，可以增强靶基因的激活。复合激活剂VPRP在不同细胞类型的一组基因中的作用比单独激活剂更有效。我们使用效应器表征了宿主染色质乙酰化和转录组的非靶标效应。我们的工作表明，转录效应和表观遗传效应可以结合使用来增强基因激活，并提示有必要进一步优化表观遗传效应以减少脱靶。
Targeted activation of endogenous genes is an important approach for cell engineering. Here, we report that the nuclease-deactivated dCas9 fused to a transcriptional activator (VPR) and an epigenetic effector (the catalytic domain of histone acetyltransferase p300(core)) simultaneously, sequentially, or as a single quadripartite effector can lead to enhanced activation of target genes. The composite activator, VPRP, behaves more efficiently than individual activators across a set of genes in different cell types. We characterize off-target effects for host chromatin acetylation and transcriptome using the effectors. Our work demonstrates that transcriptional and epigenetic effectors can be used together to enhance gene activation and suggests the need for further optimization of epigenetic effectors to reduce off-targets.