Columbia Technology Ventures

Whole methylome sequencing at the single molecule level

This technology is a method for sequencing and determining the DNA methylation profile of a genome, which can be used to screen for epigenetic biomarkers of disease.

Unmet Need: Robust and accurate genome-wide profiling of DNA methylation

DNA methylation and hydroxymethylation are epigenetic modifications of the primary sequence that impose regulatory control over the expression of various proteins. Mutations in proteins that create or process these modifications, as well as changes to the pattern can often result in diverse developmental disorders and disease. DNA methylation has traditionally been assessed with bisulfite sequencing, which is imprecise, requires large amounts of DNA, causes strand breakage, and is prone to errors in genome regions rich in cytosine-guanosine dinucleotides. Even current single molecule (SM) sequencing by synthesis and nanopore technologies cannot precisely discriminate between methylated and unmethylated DNA bases, highlighting the need for further development.

The Technology: Targeted method for detecting DNA methylation using SM sequencing

This technology uses targeted chemistry and SM sequencing for determining the DNA methylation profile of a genome. The approach overcomes the problems encountered using SM platforms by transferring bulky side groups to hydroxymethylcytosines or methylcytosines from AdoMet analogues or modified sugars to increase residence time within the nanopore. Methylated and hydroxymethylated cytosines each have a different moiety attached that would display a distinct time and magnitude signature, which can be detected in real-time using nanopore sequencing by synthesis or commercially developed SM sequencing platforms. The versatility and improved sensitivity of this technology has the potential to advance personalized medicine, providing valuable epigenetic information that can be used to diagnose and research disease.

Applications:

  • Method for epigenetic profiling
  • Research tool for studying genetic disease
  • Disease diagnostic
  • Personalized medicine
  • de novo methylation pattern identification

Advantages:

  • Simultaneously identifies different types of methylation
  • Compatible with existing SM sequencing technologies
  • More accurate and sensitive than existing methods
  • No amplification bias
  • Minimal DNA requirements allow applications in embryonic development, paraffin sections, diseased versus healthy tissues, and rare samples such as extinct species
  • Cost effective for large scale and low budget applications without sacrificing sequencing depth

Lead Inventor:

Timothy Bestor, Ph.D.

Patent Information:

Patent Status

Tech Ventures Reference: