CRISPR Guide RNA for BEST1 Allele Discrimination
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods are inadequate for effectively distinguishing and addressing dominant-mutated alleles causing genetic disorders like Best Vitelliform Macular Dystrophy, particularly in terms of knocking out the expression of disease-causing alleles while preserving functional alleles.
Innovation Solution
A method utilizing CRISPR nuclease and RNA molecules with guide sequences targeting specific SNPs in the BEST1 gene to knock out the expression of mutated alleles, allowing for the expression of functional proteins, thereby treating Best Vitelliform Macular Dystrophy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If CRISPR nuclease and guide RNA are used to target mutated alleles, then the specificity of allele discrimination is improved, but the risk of off-target effects and delivery complexity increases
Solution Approach 1:
The CRISPR system is divided into separate components (guide RNA and CRISPR nuclease) that can be independently optimized and delivered. The guide RNA contains a guide sequence portion (17-20 nucleotides) that specifically targets the mutated allele, while the CRISPR nuclease performs the cutting function. This segmentation allows for precise targeting while managing delivery complexity through separate optimization of each component.
Solution Approach 2:
The guide RNA acts as an intermediary between the CRISPR nuclease and the target DNA sequence. It contains the guide sequence portion that recognizes and binds to the mutated allele through base pairing, directing the CRISPR nuclease to the specific target site. This intermediary mechanism enables high specificity in allele discrimination while allowing the nuclease to be delivered as a separate entity.
2Reliability
If CRISPR nuclease is delivered to knock out mutated alleles, then the therapeutic effectiveness is improved, but the difficulty of in vivo delivery increases
Solution Approach 1:
The therapeutic system is segmented into deliverable components: guide RNA molecules (17-20 nucleotides) and CRISPR nuclease. This segmentation enables each component to be optimized for delivery through appropriate vectors or methods, improving the feasibility of in vivo delivery while maintaining therapeutic effectiveness through precise targeting of mutated alleles.
Solution Approach 2:
The guide RNA is designed with specific parameter optimizations (17-20 nucleotide guide sequence portion) that balance binding affinity and specificity. This parameter optimization ensures effective targeting of mutated alleles for knockout while facilitating delivery through appropriate vectors, thereby improving both therapeutic effectiveness and delivery feasibility.
3Measurement precision
If guide sequences are designed to target specific SNPs, then the precision of mutant allele identification is improved, but the guide sequence design complexity increases
Solution Approach 1:
The guide RNA contains a guide sequence portion (17-20 nucleotides) that is specifically designed to match the mutated allele sequence at the target SNP location. This local quality approach ensures high precision in SNP targeting by concentrating the specificity requirements in the guide sequence portion, while the rest of the RNA structure can be standardized for delivery and function.
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
This approach enables the specific inactivation of dominant-mutated alleles, potentially ameliorating or preventing the disease phenotype by allowing the expression of functional proteins, thus offering a targeted therapeutic strategy for genetic disorders.
Implementation Method 1
A method utilizing CRISPR nuclease and RNA molecules with guide sequences targeting specific SNPs in the BEST1 gene to knock out the expression of mutated alleles
Implementation Method 2
RNA molecules with guide sequences targeting specific SNPs in the BEST1 gene
Data Source
AI summary
RNA molecules comprising a guide sequence portion having 17-20 nucleotides in the sequence of 17-20 contiguous nucleotides set forth in any one of SEQ ID NOs: 1-3010 and compositions, methods, and uses thereof.