Fig. 1: MulTI-Tag directly identifies user-defined chromatin targets in the same cells.
From: Multifactorial profiling of epigenetic landscapes at single-cell resolution using MulTI-Tag
a, Schematic describing the MulTI-Tag methodology. (1) Antibody–oligonucleotide conjugates are used to physically associate forward-adapter barcodes with targets and are loaded directly into pA–Tn5 transposomes for sequential binding and tagmentation. (2) pA–Tn5 loaded exclusively with reverse adapters are used for a secondary CUT&Tag step to efficiently introduce the reverse adapter to conjugate-bound loci. (3) Target-specific profiles are distinguished by barcode identity in sequencing. b, Genome browser screenshot showing individual CUT&Tag profiles for H3K27me3 (first row) and RNA PolIIS5P (second row) in comparison with MulTI-Tag profiles for the same targets probed individually in different cells (third and fourth rows) or sequentially in the same cells (fifth and sixth rows). c, Heat maps describing the enrichment of H3K27me3 (red) or RNA PolIIS5P (blue) signal from sequential MulTI-Tag profiles at CUT&Tag-defined H3K27me3 peaks (left) or RNA PolIIS5P peaks (right). d, Genome browser screenshot showing H3K27me3 (red), H3K4me2 (purple) and H3K36me3 (teal) MulTI-Tag signal from experiments in H1 hESCs using an individual antibody (rows 1, 3 and 5) or all three antibodies in sequence (rows 2, 4 and 6). e, Normalized CUT&Tag (light colors) and MulTI-Tag (dark colors) enrichment of H3K27me3, H3K4me2 and H3K36me3 across genes in H1 hESCs. RPM, reads per million.
