FACT-seq

FACT-seq profiles histone modifications in formalin-fixed paraffin-embedded (FFPE) tissue samples using antibody-guided chromatin tagmentation with sequencing to enable epigenetic analysis from low-input archival specimens.

Key Features:

• High Sensitivity: Capable of profiling histone modifications from FFPE tissues with minimal input material.
• Minimal Tissue Requirement: Generates high-quality chromatin profiles from as few as ∼4000 nuclei obtained from a small FFPE tissue section.
• T7-pA-Tn5-mediated transposition: Employs a fusion protein of hyperactive Tn5 transposase and protein A (T7-pA-Tn5) for targeted chromatin tagmentation.
• T7 in vitro transcription amplification: Incorporates T7 in vitro transcription to increase sensitivity of sequencing library generation.
• Application to H3K27ac and super enhancers: Has been applied to decode H3K27ac modifications and identify disease-specific super enhancers in archived human colorectal and glioblastoma tissues.

Scientific Applications:

• Gene Regulation Studies: Enables analysis of histone modification patterns in archived FFPE samples to investigate mechanisms of gene regulation.
• Cancer Research: Facilitates identification of disease-specific epigenetic features such as super enhancers in colorectal and glioblastoma cancer tissues.
• Translational Studies: Supports epigenetic profiling in limited clinical tissue samples from archival FFPE specimens for translational research.

Methodology:

Performs antibody-guided chromatin tagmentation using T7-pA-Tn5 (hyperactive Tn5 fused to protein A) followed by T7 in vitro transcription prior to sequencing.