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. 2023 Dec 17;19(1):2293411. doi: 10.1080/15592294.2023.2293411

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© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 2. — Multifactorial CUT&Tag strategies. (left) CUT&Tag 2 for 1 involves the use of two antibodies against H3K27me3 (heterochromatin marks) and Ser5P-RNAPII (active chromatin marks). This approach follows the same workflow as regular CUT&Tag, except for mixing two antibodies into the cells. The difference in fragment size and feature breadth (broad domains for H3K27me3 and narrow peak of Ser5P-RNAPII) are used together to calculate the deconvolution of two signals. (middle) in MuTi-Tag approach, each antibody is pre-incubated with the barcoded pA-Tn5, thus marking several different histone modifications and TFs in the same experiment. (right) nano-CUT&Tag is the use of nanobodies, single-chain antibodies directly fused to Tn5, where higher efficiency can be obtained without protein A.