Multiplex Genetic Editing in Vertebrate Embryos
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Solution Overview
Problem
Current methods for making large vertebrate animals with multiple genetic changes are time-consuming and inefficient, requiring several years and multiple generations due to the serial nature of genetic editing processes, which is impractical for research and livestock production.
Innovation Solution
Simultaneous editing of multiple genes in vertebrate cells or embryos using targeted nucleases and homology-directed repair (HDR) templates, allowing for rapid creation of animals with multiple genetic edits in a single generation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If serial genetic editing is used to make multiple genetic changes, then each individual edit can be made with high precision, but the total time required increases to several years and multiple generations
Solution Approach 1:
The patent combines multiple genetic editing operations into a single simultaneous process by introducing multiple targeted nucleases and multiple HDR templates into the same cell or embryo. This merging of sequential operations into a parallel process achieves multiple genetic changes in one generation rather than requiring several years of serial breeding.
Solution Approach 2:
The patent enables continuous useful action by performing all required genetic edits in a single uninterrupted process. Instead of stopping after each edit to breed and wait for the next generation, the system continuously acts on the genome by delivering multiple editing components simultaneously, maintaining productive action throughout the process.
2Reliability
If serial breeding is used to accumulate multiple genetic changes, then each generation can be carefully selected, but the productivity decreases due to the multi-year process
Solution Approach 1:
The patent merges the functions of multiple breeding generations into a single editing event. By combining multiple genetic changes that would normally require several generations to accumulate into one simultaneous editing process, the system achieves both selection accuracy and high productivity in a single step.
Solution Approach 2:
The patent applies preliminary action by performing all necessary genetic edits before the animal is born. Instead of making edits sequentially through generations, the system prepares and delivers all editing components (nucleases and HDR templates) in advance to the embryo, ensuring all changes are made before the animal develops and can be selected.
3Productivity
If multiple targeted nucleases and HDR templates are introduced simultaneously, then multiple genes can be edited in one generation, but the process complexity increases
Solution Approach 1:
The patent applies universality by using a standardized platform that can handle multiple different nucleases and templates through common delivery mechanisms. The system uses universal components such as electroporation or microinjection for delivery, and standardized HDR template structures, allowing the same basic apparatus to perform multiple different editing operations simultaneously without requiring separate specialized equipment for each gene.
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 enables the rapid creation of animals with multiple genetic edits, reducing the time and resources needed for breeding and genetic engineering, facilitating research, livestock production, and the development of models and products for industry and medicine.
Implementation Method 1
Multiple genes can be targeted for editing using targeted nucleases and homology directed repair (HDR) templates in vertebrate cells or embryos
Data Source
AI summary
The present disclosure relates to methods for making genetic edits in vitro in a non-human vertebrate cell or embryo at a plurality of target chromosomal DNA sites. Methods for making a non-human animal having multiplex genetic edits at a plurality of target chromosomal DNA sites and making a non-human vertebrate animal chimeric for host cells and donor cells are also considered.