Conjugated CsgG-CsgF Nanopores for Clearer Nucleotide Discrimination
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Solution Overview
Problem
Existing nanopore sensing technologies for polynucleotide analytes face limitations in current range and signal-to-noise ratio (SNR), hindering effective discrimination and characterization of nucleotides.
Innovation Solution
Formation of pore complexes using CsgG pore monomers conjugated to CsgF peptides through specific linkers, such as sulfonyl fluoride, sulfonyl triazole, or fluorosulfate linkers, enhancing current range and SNR during analyte characterization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If CsgG pore monomers are used for nanopore sensing, then the pore can detect and characterize analytes, but the current range and signal-to-noise ratio are insufficient for effective nucleotide discrimination
Solution Approach 1:
The patent combines CsgG pore monomers with CsgF peptides to form hybrid pore complexes. This merging of two different protein components creates a novel structure that leverages the strengths of both proteins, resulting in improved current range and signal-to-noise ratio while maintaining analyte detection capability
Solution Approach 2:
The invention creates composite protein structures by conjugating CsgG pore monomers with CsgF peptides through specific linkers. This composite approach integrates different functional elements (the pore-forming CsgG and the signal-enhancing CsgF) into a single hybrid structure that achieves superior measurement performance
2Loss of information
If multiple nucleotides contribute to observed current, then polynucleotide translocation can be detected, but the relationship between current and nucleotide sequence becomes complex and difficult to discriminate
Solution Approach 1:
The patent introduces local modifications to the pore structure by incorporating CsgF peptides at specific locations within the pore complex. This creates localized regions with enhanced interaction properties that improve the signal contribution from individual nucleotides, making sequence discrimination more accurate despite the collective current contribution from multiple nucleotides
Data Source
AI summary
The present invention relates to novel pore monomer conjugates, pore complexes formed from the conjugates and their uses in analyte detection and characterisation.