US 12,460,246 B2
DNA-barcoded nucleosomes for chromatin mapping assays
Martis W. Cowles, Chapel Hill, NC (US); Zu-Wen Sun, Brentwood, TN (US); Michael-Christopher Keogh, Cambridge, MA (US); Bryan Jacob Venters, Cary, NC (US); and Ellen Nichole Weinzapfel, Durham, NC (US)
Assigned to EPICYPHER, INC., Durham, NC (US)
Appl. No. 17/416,928
Filed by EPICYPHER, INC., Durham, NC (US)
PCT Filed Dec. 20, 2019, PCT No. PCT/US2019/067735
§ 371(c)(1), (2) Date Jun. 21, 2021,
PCT Pub. No. WO2020/132388, PCT Pub. Date Jun. 25, 2020.
Claims priority of provisional application 62/783,861, filed on Dec. 21, 2018.
Prior Publication US 2022/0042074 A1, Feb. 10, 2022
Int. Cl. C12Q 1/6806 (2018.01); G01N 33/68 (2006.01)
CPC C12Q 1/6806 (2013.01) [G01N 33/6875 (2013.01)] 21 Claims
 
1. A nucleosome comprising: a. a protein octamer, containing two copies each of histones H2A, H2B, H3, and H4, and optionally, linker histone H1; b. a DNA molecule, comprising: i. a nucleosome positioning sequence, ii a synthetic DNA barcode indicative of the identity and/or concentration of a nucleosome feature; and iii. one or more linkers, wherein the one or more linkers comprises a nuclease and/or transposase recognition sequence; and c. a binding member linked to the DNA molecule, wherein the binding member is one half of a binding pair; wherein the nucleosome feature is none, one, or more of the histones comprising a post-translational modification or a mutation and/or a histone variant and/or the DNA molecule comprising a post-transcriptional modification wherein at least one of the one or more linkers is between the nucleosome positioning sequence and the binding member; and wherein the nucleosome is a spike-in control for a chromatin assay.