Cas13 RNA-Targeting Orthologs for Precise Eukaryotic Transcript Editing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need for affordable, easy-to-set-up, and scalable genome and transcriptome engineering technologies that can target multiple positions within eukaryotic genomes and transcriptomes, leveraging novel strategies and molecular mechanisms.
Innovation Solution
Development of RNA-targeting systems using CRISPR-Cas13 family proteins, particularly Cas13a and its orthologs, for precise manipulation of nucleic acids in eukaryotic systems, including catalytically inactive versions for targeted RNA binding and cleavage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If traditional genome editing technologies (zinc fingers, TALEs, homing meganucleases) are used, then targeted genome perturbations can be achieved, but the methods are not affordable, difficult to set up, and not scalable for targeting multiple positions
Solution Approach 1:
The patent employs CRISPR-Cas systems where a reusable Cas protein complex can be programmed with different guide RNAs to target multiple genomic positions. This copying approach allows the same effector machinery to be replicated across numerous targets through simple guide RNA design, eliminating the need to design and construct entirely new protein complexes for each target site, thereby reducing complexity while maintaining precision
Solution Approach 2:
The CRISPR-Cas system described provides universal functionality for genome editing across multiple positions. A single Cas protein system can be programmed to target any genomic location by changing only the guide RNA sequence, making the system universally applicable to multiple targets simultaneously. This multi-functionality allows the same system to perform diverse targeting tasks without requiring separate specialized tools for each position
2Manufacturing precision
If CRISPR-Cas13 family systems are used for RNA targeting, then precise manipulation of nucleic acids is achieved, but collateral activity may cause off-target effects
Solution Approach 1:
The patent extracts and utilizes the collateral RNase activity of Cas13 as a detectable signal rather than treating it purely as an off-target effect. By designing detection systems that monitor this collateral activity, the system converts a harmful byproduct into a useful diagnostic feature, allowing detection of target RNA while maintaining precision through controlled interpretation of the collateral signal
Solution Approach 2:
The system employs feedback mechanisms where collateral RNase activity is monitored and used to inform detection decisions. By measuring the level and pattern of collateral activity, the system can distinguish between specific target binding events and non-specific interactions, providing feedback that enhances targeting precision while accounting for the inherent collateral effects of Cas13
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
Enables efficient and specific RNA targeting and manipulation in eukaryotic cells, facilitating applications such as transcript knockdown, imaging, and modification of nucleic acid sequences without collateral activity.
Implementation Method 1
utilizing programmable RNA-guided RNases to achieve targeted RNA cleavage and binding
Implementation Method 2
Cas13a and Cas13b enable robust and specific RNA targeting in mammalian cells with minimal collateral activity, facilitating advanced genome editing, diagnostics, and biotechnological applications by allowing precise modification and detection of nucleic acids
Data Source
AI summary
The invention provides for systems, methods, and compositions for targeting nucleic acids. In particular, the invention provides non-naturally occurring or engineered RNA-targeting systems comprising a novel RNA-targeting CRISPR effector protein and at least one targeting nucleic acid component like a guide RNA.