High-throughput Single Cell Co-assay of Histone Modifications andTranscriptome

NIH RePORTER · NIH · R44 · $1,133,112 · view on reporter.nih.gov ↗

Abstract

Summary Disruption of gene regulation is a major causal factor in heritable disease, developmental disorders, and oncogenesis. While chromatin immunoprecipitation followed by sequencing (ChIP-seq) and Tn5- transposase based tagging (Cut&Tag) enable analysis of transcription factor binding and epigenetic state profiling in bulk tissue samples and tumor biopsies, they produce only population average signals. Yet regulatory networks and perturbations are heterogeneous between cell classes and types. Single-cell technologies can overcome the challenge of cellular heterogeneity and provide deeper insight into cell type-specific gene regulatory programs in healthy, diseased, and cancerous tissues. In prior work, we developed a single-cell joint assay of histone modification and RNA expression (Paired-Tag), enabling cell-type-stratified epigenetic profiling from bulk samples. This technology has attracted customers in both academic and nonprofit research. In the proposed study, we will develop automated protocols and effect laboratory informatics systems to establish a Paired-Tag services laboratory, we will refine our protocol to improve experimental throughput and reduce cost, and we will develop a Paired-Tag “TF” protocol for profiling transcription factor binding profiles. If successful, the research would enable next- generation multi-omic analysis of tumor or disease samples at comparable cost to single-omic technologies.

Key facts

NIH application ID
10698374
Project number
2R44GM146330-02
Recipient
EPIGENOME TECHNOLOGIES, INC.
Principal Investigator
Christopher Hartl
Activity code
R44
Funding institute
NIH
Fiscal year
2023
Award amount
$1,133,112
Award type
2
Project period
2023-07-03 → 2025-06-30