Targeted in vivo epigenome editing of H3K27me3
Targeted in vivo epigenome editing of H3K27me3
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DOI：
10.1186/s13072-019-0263-z
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发表时间：
2019-03-13
影响因子：
3.9
通讯作者：
Nakamura, Ryohei
中科院分区：
文献类型：
--
作者：
Fukushima, Hiroto S.;Takeda, Hiroyuki;Nakamura, Ryohei
背景表观遗传修饰在转录调控中起着重要作用。虽然使用下一代测序的几项研究已经揭示了表观遗传修饰和转录状态之间的全基因组关联，但由于缺乏针对表观遗传修饰的靶向操作技术，在特定基因组位点的直接因果关系尚未得到充分证明。最近，基于CRISPR-Cas9系统的表观基因组编辑技术已被报道可以直接操纵精确基因组区域的特异性修饰。然而，可编辑的修改，以及在体内应用这些技术的研究的数量仍然limited.ResultsHere，我们报告的青鳉（日本鳉，青鳉）胚胎的表观基因组的直接修改。具体地，我们开发了一种方法，使用青鳉H3 K27甲基转移酶Ezh 2（olEzh 2）和dCas 9（dCas 9-olEzh 2）的融合构建体以基因座特异性方式异位诱导阻遏性组蛋白修饰H3 K27 me 3。将dCas 9-olEzh 2 mRNA与单向导RNA（sgRNA）共注射到一细胞期胚胎中诱导H3 K27 me 3在靶位点的特异性积累并诱导基因表达下调。对活生物体中表观基因组的位点特异性操纵将导致对表观遗传修饰在发育和疾病中的作用的以前无法理解的理解。
BackgroundEpigenetic modifications have a central role in transcriptional regulation. While several studies using next-generation sequencing have revealed genome-wide associations between epigenetic modifications and transcriptional states, a direct causal relationship at specific genomic loci has not been fully demonstrated, due to a lack of technology for targeted manipulation of epigenetic modifications. Recently, epigenome editing techniques based on the CRISPR-Cas9 system have been reported to directly manipulate specific modifications at precise genomic regions. However, the number of editable modifications as well as studies applying these techniques in vivo is still limited.ResultsHere, we report direct modification of the epigenome in medaka (Japanese killifish, Oryzias latipes) embryos. Specifically, we developed a method to ectopically induce the repressive histone modification, H3K27me3 in a locus-specific manner, using a fusion construct of Oryzias latipes H3K27 methyltransferase Ezh2 (olEzh2) and dCas9 (dCas9-olEzh2). Co-injection of dCas9-olEzh2 mRNA with single guide RNAs (sgRNAs) into one-cell-stage embryos induced specific H3K27me3 accumulation at the targeted loci and induced downregulation of gene expression.ConclusionIn this study, we established the in vivo epigenome editing of H3K27me3 using medaka embryos. The locus-specific manipulation of the epigenome in living organisms will lead to a previously inaccessible understanding of the role of epigenetic modifications in development and disease.