Single Cell Chromatin Profiling by High-Efficiency Tethered Integrase

This network introduces a novel chromatin profiling technology, CUT&Tag, and will demonstrate its single cell application, thus motivating the development of new experimental and computational tools to transfer their innovation to the global Human Cell Atlas effort and the broader scientific community.

The optimization of this project’s single-cell CUT&Tag technique allows for efficient single-cell analysis of chromatin regions with specific histone modifications from complex tissues. This method has led to five publications, six protocols, three datasets, and one software pipeline, and will continue to provide insight into histone modifications and their important role in gene regulation throughout development and disease.

Papers:

• Single-cell analysis of chromatin silencing programs in developmental and tumor progression
• Profiling the epigenome at home
• CUT&Tag for efficient epigenomic profiling of small samples and single cells
• Efficient chromatin accessibility mapping in situ by nucleosome-tethered tagmentation
• Automated CUT&Tag profiling of chromatin heterogeneity in mixed-lineage leukemia

Protocols:

• Benchtop CUT&Tag, 3XFlag-pATn5 Protein Purification, and MEDS-loading
• CUT&Tag-direct with CUTAC, AutoCUT&Tag: streamlined genome-wide profiling of chromatin proteins on a liquid handling robot
• CUT&RUN: Targeted in situ genome-wide profiling with high efficiency for low cell numbers
• CUT&Tag with Drosophila tissues

Datasets:

• GSE145187, GSE158327, GSE157910