Engineered Polymerase for Orthogonal Nucleic Acid Synthesis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current technologies face challenges in producing biologically useful lengths of orthogonal nucleic acids, such as hexitol nucleic acid (HNA) and cyclohexenyl nucleic acid (CeNA), due to inefficient and expensive chemical synthesis methods, and lack of custom-made polymerases for their synthesis, replication, and evolution.
Innovation Solution
Development of nucleic acid polymerases capable of producing orthogonal nucleotides into polymers, specifically engineered to manipulate polymerase activity for HNA and CeNA, allowing for efficient transfer of genetic information and evolution of these artificial genetic systems, including the design of polymerases with mutations in key regions like the thumb region for enhanced activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If chemical synthesis methods are used to produce orthogonal nucleic acids, then production can be achieved, but the process is inefficient and expensive
Solution Approach 1:
The patent replaces chemical synthesis methods with enzymatic polymerization using engineered polymerases. This substitution of chemical processes with biological systems enables efficient production of orthogonal nucleic acids while reducing costs and improving scalability, directly resolving the contradiction between ease of manufacture and productivity
Solution Approach 2:
The engineered polymerases are designed to autonomously catalyze the polymerization of orthogonal nucleotide monomers into polymers without requiring complex chemical synthesis protocols. This self-service capability of the enzymatic system dramatically improves productivity while maintaining ease of manufacture through simple incubation conditions
2Productivity
If custom-made polymerases are developed for orthogonal nucleic acids, then synthesis and replication efficiency improves, but device complexity increases
Solution Approach 1:
The patent applies local quality by introducing specific mutations only in the thumb region (residues 651-679) of the polymerase structure, while maintaining the rest of the enzyme's native architecture. This localized modification approach enables the polymerase to handle orthogonal nucleic acids without requiring complete redesign of the entire enzyme system, thus improving productivity while limiting the increase in device complexity
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The engineered polymerases enable the efficient synthesis and replication of HNA and CeNA polymers, achieving heredity and evolution comparable to RNA virus replication, with potential applications in diagnostics, prognostics, and therapeutics, addressing systemic constraints of DNA and RNA chemistry.
Implementation Method 1
nucleic acid polymerases which are able to polymerise orthogonal nucleotides into orthogonal nucleic acid polymers
Data Source
Figure 1A~2
Figure 3
Figure 4
AI summary
The invention relates to a nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide polymer template, said polymerase comprising amino acid sequence having at least 50% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at one or more residues of the thumb region, said residues selected from: amino acids 651 to 679 (patch 10A); wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue E664, wherein said non-DNA nucleotide polymer is a RNA polymer; and wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue Y409. The invention also relates to nucleic acid encoding said enzyme, and to host cells comprising said nucleic acid.