Soluble RNA-Binding Protein with Modified Repeat Motifs
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Solution Overview
Problem
Current RNA-binding proteins lack high binding ability and specificity, particularly in recognizing target RNA sequences effectively.
Innovation Solution
Designing a soluble RNA-binding protein with modified configurations of eight repeat motifs R1 to R8 by substituting specific amino acid residues, optimizing the 13th amino acid stacking between bases to enhance binding affinity, and using extended recognition repeats to achieve high binding ability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the configuration of eight repeat motifs R1 to R8 is modified by substituting specific amino acid residues, then binding ability and specificity to target RNA sequences is improved, but protein solubility may deteriorate
Solution Approach 1:
The patent applies parameter changes by systematically substituting specific amino acid residues at defined positions (12th and 16th residues) in each of the eight repeat motifs. These parameter changes in amino acid composition enable the protein to recognize different target RNA sequences while maintaining solubility through controlled modifications rather than random changes.
Solution Approach 2:
The patent implements local quality by making targeted amino acid substitutions only at specific positions (12th and 16th residues) within each repeat motif, while keeping other regions of the protein unchanged. This localized modification approach allows the protein to gain enhanced RNA binding specificity without compromising overall protein solubility and stability.
2Reliability
If amino acid residues are substituted to optimize the 13th amino acid stacking between bases, then binding affinity is enhanced, but protein structure complexity increases
Solution Approach 1:
The patent optimizes binding affinity by changing the parameters of amino acid residues at the 13th position of each repeat motif. These parameter changes in amino acid composition enable the protein to recognize different target RNA sequences while maintaining solubility through controlled modifications rather than random changes.
3Reliability
If extended recognition repeats are used to achieve high binding ability, then binding specificity is improved, but manufacturing complexity increases
Solution Approach 1:
The patent divides the RNA recognition function into eight discrete repeat motifs (R1-R8), where each motif independently recognizes a specific nucleotide. This segmentation allows the protein to achieve high binding specificity through modular architecture, simplifying both design and manufacturing compared to creating a single large continuous recognition domain.
Solution Approach 2:
The patent creates a universal platform where the eight repeat motifs can be systematically combined and modified to recognize different target RNA sequences. By substituting amino acids at defined positions in each motif, the same basic protein structure can be adapted to bind multiple different RNA targets, reducing manufacturing complexity while maintaining high specificity.
Data Source
AI summary
An object of the present invention is to provide a soluble RNA-binding protein having high binding ability. The present invention provides an RNA-binding protein having an amino acid sequence represented by R1′-R1X—R2X—(R5X or R6Y)L-(R5X—R6Y)M-(R5X or R6Y)N—R7X—R8X—R8′ wherein each symbol means an amino acid sequence recited in the specification.