Lipid-free Anchoring of Thermophilic Bacteriophage G20C Portal Adapter into Solid-State Nanopores

A novel system for high‑resolution DNA and RNA sequencing, protein identification, and protein conformational change monitoring

Background

Nanopore-based sensors have advanced the sensitivity and selectivity of single-molecule detection. Currently, electrical and electro-optical sensors require the creation of lipid bilayers or solid-state pores in metallic membranes. Both are heavily used in biotech, but they also have limitations.This invention proposes a hybrid model where a protein pore, modeled after the G20C portal protein, is supported within a solid-state membrane. This allows for the precise engineering that comes with utilizing protein nanopores, while also providing the strength and support of a metallic membrane. These flexibilities allow this novel system to be used for high-resolution DNA and RNA sequencing, protein identification, and protein conformation change monitoring.

Technology Overview

This invention utilizes a stable and water-soluble portal protein channel, modeled after the portal protein from the thermophilic bacteriophage G20C. G20C has a robust, chemically programmable channel that can either be voltage- or pressure-inserted into a solid-state nanopore matrix to form a hybrid nanopore sensor device. Our portal protein does not require any lipid support and can be engineered to define the pore sensor’s properties. This channel can sense either electrical or optical signals, allowing the user flexibility. Optical signals provide a high-density parallel readout from multiple adjacent pores. Key elements of this innovation include mechanisms to obtain the channel’s hybrid structure, stabilize it, and modify it so that different types of biomolecules can be sensed.

Benefits

• Rapid and stable insertion of unmodified protein into a solid-state nanopore
• Mutations for sensing improvement
• Translocation of biopolymers (nucleic acids and polypeptides) for sensing applications

Applications

• High-resolution mapping of DNA, RNA sequencing, DNA sequencing
• Protein identification, protein conformational change monitoring
• Polypeptide sequencing
• Small-molecule detection, biomolecular complex detection, enzyme-ligand binding
• Biotechnology and agri-food sectors

Opportunity

• Licensing
• Research collaboration
• Sponsored research

Patents

• Utility (USA, 16/416,139)
• PCT (PCT, PCT/US2019/033006)

IP Status

• Patent application submitted
• Provisional patent

Seeking

• Development partner
• Licensing