CUT&Tag2for1: a modified method for simultaneous profiling of the accessible and silenced regulome in single cells

Cleavage Under Targets & Tagmentation (CUT&Tag) is an antibody-directed transposase-tethering chromatin profiling strategy for small samples and single cells. Previously, we showed that activation of tethered Tn5 transposase under low-salt conditions using antibodies that target promoters and enhancers produces high-resolution genome-wide chromatin accessibility maps. Here we show that low-salt CUT&Tag using a mixture of an antibody to the initiation form of RNA Polymerase II (Pol2 Serine-5 phosphate) and an antibody to repressive Polycomb domains (H3K27me3) followed by computational signal deconvolution produces efficient high-resolution maps of both the active and repressive regulomes. We have extended this CUT&Tag2for1 method to single cells using a novel deconvolution approach, thus producing high-quality multifactorial single-cell chromatin maps with a workflow that is identical to that for standard single-cell CUT&Tag. The ability to seamlessly map both the active and repressive regulatory elements in single cells provides a complete regulome profiling strategy suitable for high-throughput single-cell platforms. We used Cleavage under targets and Tagmentation (Cut-and-Tag), a chromatin profiling strategy in which antibody-targeted controlled integration of DNA sequencing adapters produces libraries for paired-end DNA sequencing.