Single-molecule regulatory architectures captured by chromatin fiber sequencing [DNAseI-seq]

We report a method for precisely stenciling the structure of individual chromatin fibers onto their composite DNA templates using non-specific DNA N6-adenine methyltransferases. Single-molecule long-read sequencing using PacBio of these chromatin stencils enables nucleotide-resolution readout of the primary architecture of multi-kilobase chromatin fibers (Fiber-seq). Overall design: Fiber-seq, AdMTase-seq and m6A-IP-seq on Drosophila S2-DRSC cells. Fiber-seq and AdMTase-seq on human K562 cells. AdMTase-seq on human Hela cells.

本研究报道了一种利用非特异性DNA N6-腺嘌呤甲基转移酶（non-specific DNA N6-adenine methyltransferases），将单个染色质纤维的结构精准复刻至其复合DNA模板之上的方法。对该染色质模板采用PacBio单分子长读长测序技术，可实现对数千碱基对染色质纤维一级结构的核苷酸分辨率读取（Fiber-seq）。整体实验设计：在果蝇S2-DRSC细胞中开展Fiber-seq、AdMTase-seq及m6A-IP-seq实验；在人类K562细胞中开展Fiber-seq及AdMTase-seq实验；在人类HeLa细胞中开展AdMTase-seq实验。