CRISPR RNA Trans-splicing for Single Sex Offspring
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Solution Overview
Problem
Current methods for distinguishing and culling male chicks in poultry are inefficient and wasteful, as they require incubation of eggs that will produce males, leading to resource waste and ethical concerns.
Innovation Solution
The use of CRISPR-Cas systems combined with RNA trans-splicing to introduce modified genes in non-human vertebrate animals, allowing for the generation of single sex offspring by making male offspring non-viable or easily identifiable.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If chicken sexers are used to determine sex of chicks, then sex identification can be performed, but it requires waiting for eggs to hatch through entire incubation process which wastes resources
Solution Approach 1:
The patent applies preliminary action by performing CRISPR-Cas9 gene editing on embryos during early development stages (in ovo or in utero) rather than waiting for hatching. This allows sex determination and selective elimination to occur before the embryo develops into a chick, thereby eliminating the waste of incubation resources while maintaining accurate sex identification through molecular methods.
2Productivity
If male chicks are culled after hatching, then single sex offspring can be achieved, but resources are wasted on incubating eggs that will produce males
Solution Approach 1:
The patent implements preliminary action by using CRISPR-Cas9 to edit embryos in ovo or in utero to eliminate unwanted sex offspring before hatching occurs. This prevents the waste of resources (food, water, incubation energy, space) that would otherwise be consumed by raising male chicks that are subsequently culled, while still achieving the goal of producing single-sex offspring populations.
Solution Approach 2:
The patent applies the extraction principle by removing the unwanted genetic material (male-determining genes or sex-specific markers) from embryos during early development through CRISPR-Cas9 editing. This allows for the selective elimination of male embryos before they develop further, thereby extracting only the desired female offspring for resource investment while eliminating the need to raise and subsequently cull male chicks.
3Productivity
If CRISPR-Cas systems with RNA trans-splicing are used, then single sex offspring can be generated by suppressing male development, but the genetic modification process becomes more complex
Solution Approach 1:
The patent uses an intermediary approach by employing RNA trans-splicing as a intermediate step in the CRISPR-Cas9 system. Instead of directly editing the genome, the system uses guide RNA that undergoes trans-splicing to form a functional complex with the Cas9 protein, which then mediates the gene editing. This intermediary RNA mechanism allows for programmable and specific gene targeting while maintaining system modularity and reducing direct manipulation of DNA.
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 efficient production of single sex populations by suppressing the development of male chicks, thereby reducing resource waste and ethical issues associated with chick culling.
Implementation Method 1
guide RNA capable of directing sequence specific binding of one or more CRISPR RNA-guided complexes encoded by the one or more expression cassettes to one or more second sequence variants of the gene
Implementation Method 2
one or more nucleotide sequences comprising a modified intron that is not capable of base pairing with a trans-splicing accepting gene
Data Source
AI summary
Described herein are methods and compositions for generating single sex offspring using enhance trans-splicing approach via an RNA binding framework. In particular, methods and compositions are provided to generate single sex and genetically modified offspring. These techniques can be applied to compassionate animal breeding.