USE OF NOVEL ERROR-CORRECTING DNA POLYMASERASES IN NEXT-GEN DNA SEQUENCING TECHNOLOGIES

Next-generation DNA sequencing has revolutionized medicine and basic research; however, current enzymes used for NGS library preparation and sequencing are prone to errors. This invention is to develop alternative NGS methods with an error-correcting enzyme

Inventor: Brian Hedlund

Dr. Brian Hedlund holds a bachelor’s degree in biology from the University of Illinois, a doctorate in microbiology from the University of Washington, and was a postdoctoral fellow at the University of Regensburg, Germany. Hedlund has published over 120 peer-reviewed scientific publications and has been a principal investigator on more than $12 million in extramural grants. 

Invention: Develop one or more thermophilic DNA polymerase enzymes (DNAP) with intrinsic uracil deglycosylase (UDG) activity for the ~$30B annual next-generation DNA sequencing (NGS) industry. Cytosine deamination, the most common mutation in nature, is the leading cause of DNA sequencing errors for all NGS platforms and lowers the sensitivity of detection of low-frequency mutations and hinders accurate genomic assemblies for mixed template applications, such as metagenomics. The proposed invention will be one or more protocols to utilize one or more natural UDG-DNAP fusion enzymes, possibly with a partner enzyme (e.g., DNA ligase), that would replace existing industry standard enzymes in NGS applications. The new UDG-DNAPs would repair naturally damaged DNA or damage incurred during library preparation and sequencing. The result would be a significant decrease in DNA sequencing errors, which would increase sensitivity for low frequency mutations and increase quality of DNA sequence data.

Benefits

• Existing NGS DNAP enzymes either have no DNA proofreading activity (e.g., Taq) or have only 3' to 5' exonuclease activity (Q5). The newly discovered UDG-DNAP fusions have 3' to 5' proofreading activity but also UDG activity. Uracil in DNA derives from the most common mutation, cytosine deamination, so the ability to remove and correct uracils would substantially improve the accuracy of NGS library preparation and sequencing.  
• NGS sequencing for diagnostics, personalized medicine, forensics, basic research.  
• Cancer diagnostics - amplification and sequencing of marker genes or whole genomes/exomes from biopsies or other samples, especially damaged samples 
• Basic research - including any application where DNA may be damaged, such as metagenomics, forensics, and ancient DNA research. 

Market Summary:

According to IBISWorld, the DNA & related laboratories in the US reach a revenue of $4.6B and are expected to grow at a CAGR of 3.4% between 2023 and 2028. Also noted on IBISWorld, the forensic technology services in the US is bringing $4.2B in revenue.

Intellectual property: 

Early stage