Chimeric Rhodopsin Gene Therapy Using Local Loop Modification
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Solution Overview
Problem
The effect of restoring visual function using ion channel type rhodopsins is insufficient, necessitating an improvement in visual function restoration and prevention of deterioration.
Innovation Solution
A chimeric protein is developed by fusing a microorganism-derived ion transport rhodopsin from the Gloeobacter genus with a bovine or human-derived G protein-coupled receptor rhodopsin, specifically modifying the second and third loops of the amino acid sequence to enhance visual function restoration and prevention of deterioration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ion channel type rhodopsins are used to restore visual function, then visual function can be restored to some extent, but the restoration effect is insufficient
Solution Approach 1:
The patent creates a chimeric protein by fusing two different rhodopsin types: the N-terminal portion (residues 1-206) from microorganism-derived ion transport rhodopsin and the C-terminal portion (residues 207-327) from animal-derived G protein-coupled receptor rhodopsin. This composite structure combines the advantages of both parent proteins to achieve superior visual function restoration compared to using either rhodopsin type alone.
Solution Approach 2:
The invention applies local quality by selectively replacing specific regions (loops 2 and 3 on the cytoplasmic side) of the microorganism-derived rhodopsin with corresponding regions from the animal-derived rhodopsin. This localized modification allows the chimeric protein to maintain the ion transport capability of the microorganism-derived portion while acquiring the G protein coupling ability of the animal-derived portion, thereby improving visual function restoration.
2Reliability
If the amino acid sequence of loops 2 and 3 is modified to improve visual function restoration, then the restoration capability is enhanced, but the protein structure becomes more complex
Solution Approach 1:
The patent modifies only specific local regions (loops 2 and 3 on the cytoplasmic side, residues 132-145 and 198-206) of the rhodopsin protein while keeping the rest of the structure intact. This localized modification approach enhances visual function restoration capability by introducing G protein coupling ability without requiring complete redesign of the entire protein structure, thus limiting the increase in complexity.
Solution Approach 2:
The invention merges the amino acid sequences of loops 2 and 3 from animal-derived G protein-coupled receptor rhodopsin into the microorganism-derived ion transport rhodopsin framework. This merging creates a chimeric protein that possesses both ion transport function and G protein coupling ability, enhancing visual function restoration while maintaining a relatively simple overall structure based on the original microorganism-derived rhodopsin.
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 chimeric protein achieves excellent visual function restoration and prevention of deterioration in conditions such as retinitis pigmentosa, age-related macular degeneration, and other retinal diseases by maintaining high activity and repeated activation.
Implementation Method 1
Rhodopsin is a photosensitive receptor with a seven transmembrane structure in the retina of humans and animals
Data Source
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AI summary
The purpose of the present invention is to provide an agent for regenerating a visual function or an agent for preventing the deterioration in a visual function, which has an excellent visual function regenerating ability. The agent for regenerating a visual function or the agent for preventing the deterioration in a visual function according to the present invention contains, as an active ingredient, a chimeric protein having both an amino acid sequence for a microorganism-origin ion-translocating receptor rhodopsin and an amino acid sequence for an animal-origin G-protein-conjugated receptor rhodopsin. The chimeric protein is preferably one in which an amino acid sequence for a cytoplasm-side second loop and/or a cytoplasm-side third loop in the amino acid sequence for the microorganism-origin ion-translocating receptor rhodopsin is substituted by an amino acid sequence for a cytoplasm-side second loop and/or a cytoplasm-side third loop in the G-protein-conjugated receptor rhodopsin.