Department of Molecular and Human Genetics, Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.
Stem Cells and Regenerative Medicine Center, Baylor College of Medicine, Houston, Texas 77030, USA.
Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026.
Comprehensive studies have shown that DNA methylation plays vital roles in both loss of pluripotency and governance of the transcriptome during embryogenesis and subsequent developmental processes. Aberrant DNA methylation patterns have been widely observed in tumorigenesis, ageing and neurodegenerative diseases, highlighting the importance of a systematic understanding of DNA methylation and the dynamic changes of methylomes during disease onset and progression. Here we describe a facile and convenient approach for efficient targeted DNA methylation by fusing inactive Cas9 (dCas9) with an engineered prokaryotic DNA methyltransferase MQ1. Our study presents a rapid and efficient strategy to achieve locus-specific cytosine modifications in the genome without obvious impact on global methylation in 24 h. Finally, we demonstrate our tool can induce targeted CpG methylation in mice by zygote microinjection, thereby demonstrating its potential utility in early development.
综合研究表明，DNA 甲基化在胚胎发生和随后的发育过程中胚胎多能性的丧失和转录组的调控中起着至关重要的作用。在肿瘤发生、衰老和神经退行性疾病中广泛观察到异常的 DNA 甲基化模式，这凸显了系统了解 DNA 甲基化以及甲基组在疾病发生和进展过程中的动态变化的重要性。在这里，我们描述了一种通过将无活性 Cas9(dCas9)与工程原核 DNA 甲基转移酶 MQ1 融合来高效靶向 DNA 甲基化的简便方法。我们的研究提出了一种快速有效的策略，可在 24 小时内实现基因组中特定基因座的胞嘧啶修饰，而对全局甲基化没有明显影响。最后，我们通过受精卵显微注射证明了我们的工具可以在小鼠中诱导靶向 CpG 甲基化，从而证明了它在早期发育中的潜在用途。
