Saltar al contenido
Merck

Single-cell CUT&Tag profiles histone modifications and transcription factors in complex tissues.

Nature biotechnology (2021-04-14)
Marek Bartosovic, Mukund Kabbe, Gonçalo Castelo-Branco
RESUMEN

In contrast to single-cell approaches for measuring gene expression and DNA accessibility, single-cell methods for analyzing histone modifications are limited by low sensitivity and throughput. Here, we combine the CUT&Tag technology, developed to measure bulk histone modifications, with droplet-based single-cell library preparation to produce high-quality single-cell data on chromatin modifications. We apply single-cell CUT&Tag (scCUT&Tag) to tens of thousands of cells of the mouse central nervous system and probe histone modifications characteristic of active promoters, enhancers and gene bodies (H3K4me3, H3K27ac and H3K36me3) and inactive regions (H3K27me3). These scCUT&Tag profiles were sufficient to determine cell identity and deconvolute regulatory principles such as promoter bivalency, spreading of H3K4me3 and promoter-enhancer connectivity. We also used scCUT&Tag to investigate the single-cell chromatin occupancy of transcription factor OLIG2 and the cohesin complex component RAD21. Our results indicate that analysis of histone modifications and transcription factor occupancy at single-cell resolution provides unique insights into epigenomic landscapes in the central nervous system.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Poli-L-lisina solution, 0.01%, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Fibronectina plasma bovino, solution, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
2- Mercaptoetanol, for molecular biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
3,3′,5-Triiodo-L-thyronine sodium salt, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Ala-Gln, 200 mM, solution, sterile-filtered, Biotechnology Performance Certified