CRISPR Gene Editing of Regulatory Elements
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Solution Overview
Problem
Current therapeutic approaches fail to effectively address the production of toxic proteins and RNA, which are implicated in various diseases such as Myotonic Dystrophy Type 1 and other neuromuscular disorders, leading to cell dysfunction and death.
Innovation Solution
The use of CRISPR gene editing to disrupt regulatory elements that drive the expression of genes associated with toxic RNA and proteins by contacting cells with guide RNAs and RNA-targeted endonucleases, specifically targeting and excising portions of genes like DMPK, to reduce the production of toxic transcripts and proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-generated harmful factors
If CRISPR gene editing is used to disrupt regulatory elements, then production of toxic proteins and RNA is reduced, but risk of off-target effects and genomic instability increases
Solution Approach 1:
The patent applies local quality by designing guide RNAs that target specific regulatory elements (promoters, enhancers, introns) with high precision. The CRISPR-Cas system is configured to modify only specific genomic loci containing disease-causing sequences, leaving the rest of the genome unchanged. This localized approach reduces off-target effects while effectively disrupting toxic transcript production at the targeted regulatory regions.
Solution Approach 2:
The patent employs partial action by using CRISPR to disrupt only the regulatory elements necessary to reduce toxic protein and RNA production, rather than eliminating entire genes. This partial disruption achieves therapeutic effect while minimizing genomic changes and potential off-target consequences. The approach modifies specific cis-regulatory sequences without causing excessive genomic instability.
2Object-generated harmful factors
If regulatory elements are disrupted to reduce toxic transcript production, then disease phenotype is ameliorated, but normal gene expression control may be affected
Solution Approach 1:
The patent applies local quality by designing guide RNAs that target specific regulatory elements (promoters, enhancers, introns) with high precision. The CRISPR-Cas system is configured to modify only specific genomic loci containing disease-causing sequences, leaving the rest of the genome unchanged. This localized approach reduces off-target effects while effectively disrupting toxic transcript production at the targeted regulatory regions.
Solution Approach 2:
The patent converts the harmful expanded repeat sequences and aberrant regulatory elements into a therapeutic opportunity. By targeting these disease-causing sequences with CRISPR, the system transforms the problematic regulatory regions into the very target of treatment. The disrupted regulatory elements that originally caused toxic transcript production now serve as the focal point for reducing pathogenic expression, turning the disease mechanism into the solution.
3Ease of manufacture
If CRISPR components are delivered to cells, then gene editing can be performed, but delivery complexity and cell toxicity increase
Solution Approach 1:
The patent applies segmentation by dividing the CRISPR delivery system into separate modular components: guide RNAs, Cas proteins, and delivery vehicles (viral or non-viral vectors). Each component can be optimized and delivered independently, allowing for tailored approaches to different cell types and disease models. This modular segmentation reduces overall delivery complexity compared to attempting to deliver a complete, integrated editing system.
Solution Approach 2:
The patent employs intermediary delivery vehicles (viral vectors such as AAV, or non-viral liposomes and electroporation reagents) that mediate the transfer of CRISPR components into cells. These intermediaries protect the delicate guide RNA and Cas protein from degradation during delivery, facilitate cellular uptake, and reduce direct toxicity. The delivery vehicle acts as a buffer and transport mechanism, simplifying the overall delivery process while maintaining cell viability.
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 method effectively reduces the production of toxic proteins and RNA, thereby ameliorating disease phenotypes by targeting and excising specific regulatory elements, as demonstrated in Myotonic Dystrophy Type 1 and potentially other disorders.
Implementation Method 1
the guide RNA comprises a guide sequence that directs the RNA-targeted endonuclease to or near one or more regulatory elements
Implementation Method 2
an RNA-targeted endonuclease, or a nucleic acid encoding the RNA-targeted endonuclease, wherein the guide RNA comprises a guide sequence that directs the RNA-targeted endonuclease to or near one or more regulatory elements
Data Source
AI summary
Compositions and methods for disrupting and/or excising regulatory elements or portions thereof that are associated with genes that produce toxic proteins and transcripts are encompassed.