Dam&ChIC v1 Article Swipe
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· 2025
· Open Access
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· DOI: https://doi.org/10.17504/protocols.io.261gee4njg47/v1
· OA: W4411836230
Dam&ChIC is a single-cell chromatin profiling method that allows for multifactorial and retrospective measurements of chromatin factors. It builds-upon and integrates scDamID and sortChIC, which are both methods for profiling of chromatin factors at high resolution in single cells. Dam&ChIC combines the properties of its predecessor approaches, which are fundamentally different. In DamID chromatin states are recorded over time in living cells, due to the capacity of Dam to methylate the chromatin in its proximity with a stable exogenous to eukaryotes modification (m6A). On the other hand, ChIC captures the most recent chromatin state in fixed cells using antibody-mediated targeting of pA-MNase on the chromatin. Thus, Dam&ChIC provides a cumulative past readout by its DamID modality, coupled to a present-state readout by its ChIC modality. As such, the method can be used to (i) profile two different chromatin factors in single cells to dissect their interplay, or (ii) profile the same chromatin factor retrospectively and obtain insights into its dynamics over time. Here we describe the workflow of single-cell Dam&ChIC, including a strategy for fluorescence-based sample multiplexing ("hashing") with CellTrace dyes, that enables parallel processing of cells derived from multiple experimental conditions.
