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DMF-ChIP-seq用于低输入细胞的高度敏感和集成的表观基因组分析

Mingyin Li¹, Xing Na¹, Fanghe Lin¹

¹The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, the Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, Department of Chemical Biology, Department of Chemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.

ACS applied materials & interfaces

|September 20, 2024

概括

我们开发了DMF-ChIP-seq,这是一种数字微流体方法,用于对全基因组的DNA-蛋白相互作用 (DPI) 进行分析. 这种技术提供了高灵敏度和效率,需要最小的细胞进行精确的表观基因组分析.

科学领域:

表观遗传学和基因组学
生物技术是生物技术.
分子生物学分子生物学

背景情况:

全基因组的DNA-蛋白相互作用 (DPI) 对于理解表观遗传调节至关重要.
目前的DPI分析方法面临诸如高细胞要求,低灵敏度和大量试剂使用等挑战.

研究的目的:

开发一种新,高效和用户友好的基因组范围DPI分析方法.
克服现有的DPI分析技术的局限性.

主要方法:

开发DMF-ChIP-seq,在单个数字微流体芯片上集成细胞预处理,抗体识别,标记,丰富和PCR放大.
利用封闭的亚微升反应体积和超疏水界面来提高反应效率.

主要成果:

与当前方法相比,DMF-ChIP-seq在峰值缩中表现出更高的灵敏度.
实现了高精度 (PCC>0.86) 和可重复性 (PCC>0.92).
该方法成功地从8个单元中对DPI进行了分析,其信号噪声比高.

结论:

DMF-ChIP-seq为全基因组DPI分析提供了一种高效,敏感和具有成本效益的方法.
这项技术可以对表观基因组景观动态进行详细的研究,例如早期胚胎分化期间的H3K27ac修饰.
该系统对研究各种生物过程中的表观遗传调节具有重大前景.

关键词:

DNA - 蛋白相互作用数字微流体学数字微流体学表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.基因组胺基因的修改测序测序是指测序的时间.

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相关概念视频

Chromatin Immunoprecipitation- ChIP

Chromatin Immunoprecipitation- ChIP

Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...