Escorted Guide RNA for CRISPR Spatial Control
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Solution Overview
Problem
Current CRISPR-Cas systems lack spatial and temporal control over gene editing, leading to off-target effects and undesired sustained expression, which can cause pathological conditions due to dysregulation of gene expression.
Innovation Solution
Development of escorted CRISPR-Cas systems with modified guide RNAs (esgRNAs) that include an escort RNA aptamer sequence for targeted delivery and activation/deactivation based on specific cellular conditions, such as light exposure or pH, allowing for precise temporal and spatial control of gene editing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If CRISPR-Cas systems are used for gene editing, then gene editing efficiency is improved, but off-target effects and unintended genomic alterations occur due to lack of spatial and temporal control
Solution Approach 1:
The guide RNA is modified with an escort aptamer sequence beforehand, which enables targeted delivery to specific cellular locations before the CRISPR-Cas system becomes active. This preliminary functionalization ensures that the system only acts where intended, preventing off-target effects while maintaining editing efficiency
Solution Approach 2:
The escort RNA aptamer sequence acts as an intermediary between the guide RNA and the target cellular component (such as a cell surface protein or intracellular marker). This intermediary enables precise spatial control by mediating the delivery of the CRISPR-Cas system to the correct location, thereby reducing unintended genomic alterations
2Duration of action of stationary object
If CRISPR-Cas systems are used for sustained gene expression control, then gene expression regulation is improved, but pathological conditions may occur due to dysregulation of gene expression
Solution Approach 1:
The CRISPR-Cas system is designed with dynamic temporal control through the escort aptamer mechanism, allowing the system to be activated only when and where the aptamer binds to its target. This dynamic control enables sustained gene expression regulation at the intended location and time, preventing dysregulation and pathological conditions
Solution Approach 2:
The system utilizes changes in cellular parameters (such as pH, presence of specific proteins, or other localized conditions) to trigger aptamer binding and activate CRISPR-Cas activity. This parameter-based control ensures that gene expression is regulated accurately according to the desired temporal and spatial parameters, maintaining reliability
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
Enhances the specificity and efficiency of gene editing by ensuring that CRISPR activity is restricted to intended genomic loci, reducing off-target effects and enabling time-limited expression to prevent unintended genomic alterations.
Implementation Method 1
the escort aptamer (i.e. the escort RNA aptamer sequence) has binding affinity for an aptamer ligand on or in the cell
Implementation Method 2
The aptamer effector may be a transient effector, such as an external energy source that is applied to the cell at a particular time. In some embodiments, the external energy source is light energy
Data Source
AI summary
The present invention generally relates to CRISPR systems or complexes, such as those with an escorted guide RNA.