Mitochondrial Base Editing for Precise LHON Mutation Correction
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Solution Overview
Problem
There is no effective treatment for Leber hereditary optic neuropathy (LHON), a mitochondrial genetic disorder caused by specific mutations in mitochondrial DNA, leading to irreversible vision loss, with current treatments only delaying progression without curing the disease.
Innovation Solution
A base editing system using fusion proteins or polynucleotides encoding them, comprising DNA binding proteins and deaminases, specifically targets and corrects adenine to guanine at position 3460, adenine to guanine at position 11778, or cytosine to thymine at position 14484 in mitochondrial DNA to revert mutations to a normal genotype.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If base editing system is used to correct mitochondrial DNA mutations, then treatment effectiveness is improved, but device complexity increases
Solution Approach 1:
The base editing system is divided into separate functional modules: a mitochondrial DNA-targeting module (using mtDNA-specific gRNA and DNA binding protein), a base editing module (using deaminase enzyme), and a delivery module. This segmentation allows each component to be optimized independently while maintaining overall system effectiveness for correcting LHON mutations.
Solution Approach 2:
The patent introduces intermediary components such as mitochondrial targeting sequences (MTS) that mediate the delivery of base editing components to mitochondrial DNA, and uses specific DNA binding proteins as intermediaries between the guide RNA and the target mitochondrial genome. These intermediaries enable precise targeting while simplifying the overall system architecture.
2Manufacturing precision
If specific base editing is performed to correct mutations, then manufacturing precision is improved, but device complexity increases
Solution Approach 1:
The base editing system employs local quality by using mitochondrial DNA-specific guide RNAs that target only the mutated bases at positions 3460, 11778, or 14484 in the mitochondrial genome. The deaminase enzyme is localized to act only at these specific target sites through the guidance of mtDNA-specific sequences, ensuring high editing precision while maintaining a relatively simple overall structure.
Solution Approach 2:
The system achieves precise base editing by changing specific parameters: using mtDNA-specific gRNA sequences with unique mitochondrial promoters and origin of replication sequences, adjusting the deaminase enzyme's activity parameters, and controlling the editing conditions to achieve high precision correction of only the intended mitochondrial mutations without affecting nuclear DNA.
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 system effectively prevents or treats LHON by correcting the underlying genetic mutations, potentially restoring normal function and preventing vision loss.
Implementation Method 1
an activity of specifically correcting the adenine base at position 3460 or 11778, or the cytosine base at position 14484
Data Source
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AI summary
The present invention relates to a base editing system for correcting mutations G3460A, G11778A, or T14484C in mitochondrial DNA of a patient with Leber hereditary optic neuropathy (LHON) to a normal genotype. Specifically, the present invention provides a method for correcting a mutation in the mitochondrial genes of a patient with LHON to a normal genotype using a base editor that recognizes specific sites in the mitochondrial genes of the patient with LHON and has an activity of specifically correcting the adenine base at position 3460 or 11778, or the cytosine base at position 14484, by using a fusion protein or a polynucleotide encoding such a fusion protein. The base editor or nucleotide according to the invention may correct DNA mutations specific to LHON in a cellular or extracellular in vitro environment and, more preferably, may be used as a gene therapy agent for the prevention or treatment of the disease. Thus, the present invention also provides a use of the substance in the prevention or treatment of LHON.