On-Substrate Nucleic Acid Synthesis With Sequencing Feedback
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Solution Overview
Problem
Existing nucleic acid synthesis techniques are labor-intensive, time-consuming, and prone to errors, especially when synthesizing long polynucleotide chains, and lack efficient quality control for creating entirely synthetic, template-independent molecules.
Innovation Solution
The techniques incorporate quality feedback through sequencing data during synthesis phases, allowing for the selection and assembly of nucleic acid fragments with desired quality, and utilize a single platform for simultaneous synthesis and sequencing to enhance accuracy and efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Length of moving object
If chemical solid phase synthesis is used to form long polynucleotide chains, then the synthesis capability is achieved, but the error rate increases significantly
Solution Approach 1:
The patent divides the synthesis process into multiple cycles where oligonucleotides are synthesized in segments, quality-controlled through sequencing, and then assembled into longer polynucleotide chains. This segmentation allows each segment to be verified for accuracy before being incorporated into the final product, thereby maintaining high sequence accuracy while achieving long chain lengths.
Solution Approach 2:
The patent implements quality feedback by performing sequencing on synthesized oligonucleotides during the synthesis process. This feedback mechanism identifies sequences with errors, allowing only high-quality sequences to be selected and assembled into the final polynucleotide product, thus resolving the contradiction between chain length and sequence accuracy.
2Ease of manufacture
If traditional recombinant techniques are used to introduce mutations, then mutation generation is achieved, but labor-intensive analysis and time-consuming host cell propagation are required
Solution Approach 1:
The patent replaces traditional mechanical and biological methods (host cell propagation) with chemical synthesis and sequencing-based quality control. By using chemical solid phase synthesis combined with sequencing feedback to identify and select correct sequences, the patent eliminates the need for time-consuming host cell propagation while maintaining the ability to generate desired mutations.
3Manufacturing precision
If chemical solid phase synthesis is used for short sequences, then synthesis accuracy is maintained, but synthesis speed decreases for long sequences
Solution Approach 1:
The patent synthesizes oligonucleotides in manageable segments using chemical solid phase synthesis, then assembles these verified segments into longer polynucleotide chains. This segmentation maintains synthesis accuracy for each segment while improving overall productivity by parallelizing the synthesis and assembly process.
Solution Approach 2:
The patent performs quality control through sequencing on synthesized oligonucleotides before assembly into final products. This preliminary action ensures that only accurate sequences are selected and assembled, maintaining high manufacturing precision while streamlining the overall process to improve productivity.
Data Source
AI summary
A method for synthesizing a nucleic acid includes synthesizing one or more nucleic acid fragments on a substrate. The synthesized one or more nucleic acid fragments may be amplified on the substrate. The method also includes sequencing the synthesized or amplified one or more nucleic acid fragments on the substrate. The sequencing may provide feedback to designs of the one or more nucleic acid fragments. The method further includes harvesting the synthesized or amplified one or more nucleic acid fragments based on sequencing. The synthesized or amplified one or more nucleic acid fragments may be assembled to generate a target nucleic acid.