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通过SelectID在甲基化基因组位点上对表观遗传调控因子进行选择性鉴定。

Selective identification of epigenetic regulators at methylated genomic sites by SelectID.

作者信息

Qian Wenchang, Jiang Penglei, Niu Mingming, Fu Yujuan, Huang Deyu, Zhang Dong, Liang Ying, Wang Qiwei, Han Yingli, Zeng Xin, Shi Yixin, Jiang Lingli, Yu Zebin, Li Jinxin, Lu Huan, Wang Hong, Chen Baohui, Qian Pengxu

机构信息

Center for Stem Cell and Regenerative Medicine and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, State Key Laboratory of Experimental Hematology, Hangzhou, China.

Liangzhu Laboratory, Zhejiang University, 1369 West Wenyi Road, Hangzhou, China.

出版信息

Nat Commun. 2025 Apr 18;16(1):3709. doi: 10.1038/s41467-025-59002-y.

Abstract

DNA methylation is a significant component in proximal chromatin regulation and plays crucial roles in regulating gene expression and maintaining the repressive state of retrotransposon elements. However, accurate profiling of the proteomics which simultaneously identifies specific DNA sequences and their associated epigenetic modifications remains a challenge. Here, we report a strategy termed SelectID (selective profiling of epigenetic control at genome targets identified by dCas9), which introduces methylated DNA binding domain into dCas9-mediated proximity labeling system to enable in situ protein capture at repetitive elements with 5-methylcytosine (5mC) modifications. SelectID is demonstrated as feasible as dCas9-TurboID system at specific DNA methylation regions, such as the chromosome 9 satellite. Using SelectID, we successfully identify CHD4 as potential repressors of methylated long interspersed nuclear element-1 (LINE-1) retrotransposon through direct binding at the 5' untranslated region (5'UTR) of young LINE-1 elements. Overall, our SelectID approach has opened up avenues for uncovering potential regulators of specific DNA regions with DNA methylation, which will greatly facilitate future studies on epigenetic regulation.

摘要

DNA甲基化是近端染色质调控的重要组成部分,在调节基因表达和维持逆转录转座子元件的抑制状态中发挥关键作用。然而,同时识别特定DNA序列及其相关表观遗传修饰的蛋白质组学精确分析仍然是一项挑战。在此,我们报告了一种称为SelectID(通过dCas9鉴定的基因组靶点处表观遗传控制的选择性分析)的策略,该策略将甲基化DNA结合结构域引入dCas9介导的邻近标记系统,以实现对具有5-甲基胞嘧啶(5mC)修饰的重复元件进行原位蛋白质捕获。在特定的DNA甲基化区域,如9号染色体卫星区域,SelectID被证明与dCas9-TurboID系统一样可行。使用SelectID,我们通过直接结合年轻的LINE-1元件的5'非翻译区(5'UTR),成功鉴定出CHD4作为甲基化长散在核元件-1(LINE-1)逆转录转座子的潜在抑制因子。总体而言,我们的SelectID方法为揭示具有DNA甲基化的特定DNA区域的潜在调节因子开辟了道路,这将极大地促进未来的表观遗传调控研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6521/12008204/68268cd82510/41467_2025_59002_Fig1_HTML.jpg

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