Fucntional genetic variants mediate their regulatory effects through altered transcription factor binding. [ATAC-Seq]

The extensive molecular characterization of the transcriptional regulatory landscape within the LCLs of a 3-generation, 17-member kindred was performed to allow for the identification of functional variants. A variant associated with changes in multiple molecular phenotypes was selected for validation through precise CRISPR/Cas9 editing. Edited clones were subsequently tested for reconstitution of function through CRISPR/dCas9 targeted TF activation. Overall design: The accessible regions of chromatin from a 3-generation,17-member kindred were assayed from two different cultures; thus, producing two biological replicates.

本研究对一个包含三代共17名成员的家系的淋巴母细胞样细胞系（Lymphoblastoid Cell Lines, LCLs）内的转录调控图谱开展了全面的分子表征，以实现功能性变异的鉴定。本研究选取了与多种分子表型改变相关的变异，通过精准的CRISPR/Cas9基因编辑对其进行验证。随后，通过CRISPR/dCas9靶向转录因子（Transcription Factor, TF）激活实验，对编辑后的克隆株的功能重建情况进行了检测。
整体实验设计：该家系的染色质开放区域通过两种不同的培养体系进行检测，由此得到两份生物学重复样本。