CUTANA™ Hia5 for Fiber-seq

CUTANA™ Hia5 is the key enzyme for Fiber-seq [1], the multiomic long-read sequencing (LRS) technique that simultaneously profiles chromatin accessibility, DNA methylation, and genetic variation at single-molecule resolution.

Benefits

• Multiomic insights in one assay – Fiber-seq simultaneously profiles chromatin accessibility, DNA methylation, and genetic variants, consolidating what typically requires three or more separate assays into one long-read sequencing assay.
• Highest activity 6mA-MTase – Faster labeling via Hia5 enables higher resolution of accessible DNA, with demonstrated utility in protein footprinting applications to resolve transcription factor binding and nucleosome positioning.
• Access complex genomic regions – Profile chromatin accessibility in repetitive and structurally complex regions such as transposable elements, centromeres, telomeres, segmental duplications, and chromosomal rearrangements.
• Easy workflow integration – Fiber-seq can be performed in <2 hours directly upstream of routine long-read sequencing library preparation.
• Resolve chromatin state heterogeneity – PCR-free approach leverages long-read sequencing to directly profile individual DNA molecules, uncovering chromatin structure heterogeneity in cell populations often missed by short-read techniques.
  
  

Questions?

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Function of Hia5 Enzyme

Hia5 is a DNA adenine methyltransferase (MTase) that uses S-adenosylmethionine (SAM) to catalyze methylation at the N6 position of adenine, producing N6-methyladenine (6mA). Fiber-seq leverages the nonspecific activity of Hia5 to methylate adenines within accessible chromatin regions in situ. This records chromatin accessibility information onto native DNA while preserving endogenous methylation (5mC). Hia5-modified DNA is fully compatible with LRS on both Pacific Biosciences® (PacBio® HiFi Sequencing) and Oxford Nanopore Technologies® (ONT® Nanopore Sequencing) platforms.

Workflow

By layering chromatin accessibility on top of standard LRS workflows, Fiber-seq uniquely links epigenetic information (DNA methylation) and complex genetic architecture (e.g., structural variants and haplotype phasing) to chromatin states within the same molecules of DNA.

Figure 1: Fiber-seq methods.

Performance Data

Figure 2: Fiber-seq simultaneously detects open chromatin (6mA) and DNA methylation (5mC)

Figure 3: In vitro confirmation of Hia5 6mA methyltransferase activity

Figure 4: SDS-PAGE analysis of Hia5 purity

Any Questions Left?

Contact us for assistance to get started on your Fiber-seq sequencing experiment.

Want to learn more about Fiber-seq?

View the Science publication Single-molecule regulatory architectures captured by chromatin fiber sequencing, A.B.Stergachis, et al. (2020).