Single-molecule regulatory architectures captured by chromatin fiber sequencing [Hs Fiber-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.