Sequential Gene Editing Method for Reducing Translocation Risks
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Solution Overview
Problem
CRISPR/Cas systems for gene editing pose risks such as off-target effects, translocation events, and potential malignancy, particularly when making multiple genetic modifications in cells.
Innovation Solution
The method involves sequentially introducing genetic modifications to cells, ensuring that each break in the genomic DNA is substantially repaired before introducing the next break, thereby minimizing the production of translocation products.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple genetic modifications are introduced simultaneously using CRISPR/Cas systems, then the productivity of gene editing is improved, but the risk of translocation events and off-target effects increases
Solution Approach 1:
The patent divides the simultaneous introduction of multiple genetic modifications into sequential steps. Instead of introducing all gRNAs and nucleases at once, the method introduces them in separate rounds with time intervals, allowing DNA breaks to be repaired between introductions. This segmentation reduces the probability of translocation events while maintaining cumulative editing efficiency.
Solution Approach 2:
The patent applies preliminary action by allowing the first DNA break to be substantially repaired before introducing the second break. This is achieved by introducing the first gRNA and nuclease, waiting for repair (which can be monitored), and then introducing subsequent editing components. This preliminary repair action prevents harmful translocation events.
2Reliability
If multiple genetic modifications are introduced sequentially with time intervals, then the safety regarding translocation events is improved, but the duration of the gene editing process increases
Solution Approach 1:
The patent employs periodic action by introducing editing components in discrete rounds separated by specific time intervals. Each round consists of introducing gRNA and nuclease, allowing a defined period for repair, then repeating the process. This periodic approach balances safety (by allowing repair) with efficiency (by maintaining a rhythm of editing activities).
Solution Approach 2:
The patent changes the temporal parameters of the editing process by optimizing the time intervals between introductions. By adjusting these parameters (time intervals, concentration, timing), the method achieves the shortest safe duration that still allows adequate repair, thus minimizing time loss while maintaining safety.
3Adaptability or versatility
If CRISPR/Cas systems are used for therapeutic applications, then the versatility of gene editing is improved, but the potential malignancy risk increases
Solution Approach 1:
The patent applies preliminary anti-action by proactively preventing translocation events and off-target effects before they can lead to malignancy. The sequential introduction method with repair intervals serves as a preventive measure against the harmful effects that could otherwise arise from the versatile CRISPR/Cas system, thereby counteracting potential malignancy risk.
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 approach reduces the risk of generating translocation products, enhancing the safety and efficacy of multiple genetic modifications in therapeutic applications.
Implementation Method 1
Clustered regulatory Interspaced Short Palindromic Repeats (CRISPR)/Cas systems provide a platform for targeted gene editing in cells
Implementation Method 2
breaks in genomic DNA can be recognized and repaired by a variety of cellular DNA repair processes
Data Source
AI summary
Provided herein are methods of making genetically engineered cells having a plurality of modifications (e.g., insertions or deletions), cells and cell populations produced by said methods, methods involving administering such genetically engineered cells to a subject, such as a subject having a hematopoietic malignancy.