Zinc Finger Proteins Modulate Htt Allele Expression
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Solution Overview
Problem
Current treatments for Huntington's Disease are limited, and there is a need for effective methods and compositions to modify the expression of the Huntingtin (Htt) allele to treat and prevent the disease.
Innovation Solution
Engineered DNA binding domains, such as zinc finger proteins and TALE proteins, are used to modulate the expression of the HD Htt allele by binding to specific sequences, either repressing or activating the mutant or wild-type alleles, and CRISPR/Cas systems are employed to target and modify the genomic structure of the Htt gene.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current treatments are used for Huntington's Disease, then disease progression is managed, but effective modification of Htt allele expression is not achieved
Solution Approach 1:
The patent extracts and targets the specific CAG repeat region within the Htt gene using engineered DNA binding domains. By focusing the therapeutic action on the problematic expanded repeat sequence rather than the entire gene, the treatment aims to selectively modify mutant allele expression while preserving wild-type function, thereby improving treatment effectiveness without compromising disease progression control
Solution Approach 2:
The invention applies local quality by designing DNA binding domains with specific affinity for the expanded CAG repeat sequences. The binding domains are engineered to recognize and bind preferentially to the abnormal repeat structure, enabling localized modulation of gene expression at the precise site of pathology while leaving the rest of the gene and genome unaffected
2Reliability
If zinc finger proteins and TALE proteins are used to modulate Htt allele expression, then mutant allele repression is achieved, but device complexity increases
Solution Approach 1:
The patent segments the DNA binding function into modular zinc finger or TALE repeat units, where each unit recognizes a specific triplet or dinucleotide sequence. This segmentation allows the binding domain to be customized for different target sequences by simply changing the number and type of modular units, reducing the overall complexity compared to designing entirely novel proteins for each target
Solution Approach 2:
The invention employs universal DNA binding platforms (zinc finger proteins and TALE proteins) that can be adapted to recognize various DNA sequences through modular assembly. These platforms serve multiple functions: sequence-specific binding, dimerization for enhanced affinity, and potential fusion to transcriptional regulatory domains, thereby achieving mutant allele repression with a versatile toolset rather than requiring separate custom-designed molecules for each application
3Adaptability or versatility
If CRISPR/Cas systems are employed to modify Htt gene genomic structure, then gene editing capability is improved, but difficulty of detecting and measuring increases
Solution Approach 1:
The patent introduces intermediary reporter systems and molecular beacons that facilitate detection of CRISPR/Cas-mediated genomic modifications. These intermediaries include fluorescently labeled probes that bind to specific genomic loci, allowing visualization and quantification of editing outcomes without requiring complex genomic sequencing or PCR-based methods
Data Source
AI summary
Disclosed herein are compositions comprising non-naturally occurring zinc finger domains, fusion proteins comprising these zinc finger domains, polynucleotides encoding these proteins, cells expressing these proteins and pharmaceutical compositions comprising these proteins or polynucleotides as well as methods of modifying an Htt gene using these compositions for treating or preventing Huntington's Disease.