Modified Nucleotide Analogues for Accurate Sequencing by Synthesis
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Solution Overview
Problem
Existing DNA sequencing methods face challenges in efficiently and accurately incorporating modified nucleotides and nucleosides as substrates during sequencing by synthesis (SBS) to achieve the goal of the $1,000 genome.
Innovation Solution
Development of novel nucleotide analogues with specific functional groups that are recognized by DNA polymerases, allowing for accurate incorporation into growing DNA chains and subsequent sequencing through detectable labeling.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional nucleotides are used in sequencing by synthesis, then the sequencing process can proceed with standard DNA polymerases, but the accuracy and efficiency of nucleotide incorporation is limited
Solution Approach 1:
The patent modifies the chemical structure of nucleotides by changing parameters such as adding polyphosphate moieties, nucleoside protecting groups, and polymerase-compatible cleavable moieties. These structural parameter changes enable conventional DNA polymerases to recognize and incorporate the modified nucleotides with high accuracy and efficiency, thereby improving sequencing performance without requiring new polymerase enzymes.
2Reliability
If modified nucleotides with complex structures are developed, then recognition and incorporation by polymerases is improved, but the complexity of nucleotide synthesis and characterization increases
Solution Approach 1:
The patent divides the modified nucleotide into distinct functional segments: the nucleobase, the sugar moiety, the polyphosphate group, the protecting group, and the cleavable moiety. Each segment has a specific function, and this segmentation allows for modular design and simplifies the synthesis and characterization processes while maintaining reliable polymerase recognition and incorporation.
3Adaptability or versatility
If multiple types of modified nucleotides are synthesized and tested, then the versatility of sequencing applications is improved, but the time and resources required for synthesis and characterization increase
Solution Approach 1:
The patent creates a universal platform of modified nucleotides that can be applied to multiple sequencing applications. By developing nucleotides with different combinations of polyphosphate groups, protecting groups, and cleavable moieties, the same basic nucleotide structure can serve various sequencing purposes, reducing the need to synthesize and characterize entirely new nucleotides for each application.
Data Source
AI summary
Disclosed herein, inter alia, are compounds, modified nucleotides, compositions, and methods of using the same.