Cas13 RNP Targeting Nucleotide Repeat Expansions

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Solution Overview

Problem

Current treatments for diseases caused by nucleotide repeat expansions, such as amyotrophic lateral sclerosis and frontotemporal dementia, are ineffective due to the difficulty in targeting these expansions with conventional technologies.

Innovation Solution

A ribonucleoprotein (RNP) comprising a Cas13 protein and a guide RNA (gRNA) with a spacer sequence containing CCGGCC or CCCCGG sequences is used to specifically target and reduce the expression of toxic nucleotide repeat expansions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional technologies are used to target nucleotide repeat expansions, then treatment effectiveness is poor, but the difficulty in targeting these expansions remains high

Engineering Contradiction:

Improvetreatment effectivenessVSAvoiddifficulty in targeting nucleotide repeat expansions

Core Design Contradiction:

ReliabilityVSDifficulty of detecting and measuring

Solution Approach 1:

The patent employs guide RNA molecules as intermediaries that specifically bind to nucleotide repeat expansion sequences, enabling the Cas13 protein to accurately target and cleave toxic RNA transcripts. This intermediary mechanism overcomes the difficulty of directly targeting repetitive sequences with conventional technologies.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent replaces conventional mechanical or chemical targeting methods with a biological recognition system. The guide RNA uses base-pairing complementarity to recognize and bind to specific nucleotide repeat sequences, while the Cas13 protein provides enzymatic cleavage activity, substituting physical targeting approaches with molecular recognition and catalysis.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Object-generated harmful factors

If Cas13 protein and gRNA are used to target repeat-containing RNA, then toxic RNA and protein levels are reduced, but device complexity increases

Engineering Contradiction:

Improvetoxic RNA and protein levelsVSAvoidRNP complex structure

Core Design Contradiction:

Object-generated harmful factorsVSDevice complexity

Solution Approach 1:

The patent divides the targeting system into two functional segments: the Cas13 protein that provides catalytic activity and the guide RNA that provides sequence-specific recognition. This segmentation allows each component to be optimized independently and simplifies the overall design compared to a single complex molecule.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The Cas13 protein serves multiple functions: it binds to the guide RNA, recognizes the target sequence through the guide RNA, and cleaves the toxic RNA transcript. This multi-functionality reduces the need for additional separate components, thereby managing complexity while achieving effective toxin reduction.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 RNP effectively decreases the levels of toxic RNA and protein products generated from nucleotide repeat expansions, providing a potential therapeutic approach for neurological diseases associated with these expansions.

Implementation Method 1

a ribonucleoprotein (RNP) comprising a Cas13 protein and a gRNA, wherein said gRNA comprises a spacer sequence comprising one or more CCGGCC sequences

Methodology Applied

Scientific EffectCRISPR-Cas13 RNA-guided RNA targeting:

Data Source

PatentUS20250154480A1Treatment for nucleotide repeat expansion disease

Publication Date: 2025.05.15 JOHNS HOPKINS UNIVERSITY

AI summary

Provided herein is technology relating to nucleotide repeat elements present in the genomes of eukaryotes and particularly, but not exclusively, to technologies for reducing the levels of disease-causing products expressed from expansions of nucleotide repeats.