CRISPR-Cas9 Editing of MS1 and MS5 Genes for Male Sterility
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Solution Overview
Problem
Current methods for inducing male sterility in plants, such as chemical or random mutagenesis, are inefficient and require extensive screening and backcrossing to eliminate undesired mutations, making them labor-intensive and prone to errors.
Innovation Solution
Introducing genetic modifications into specific endogenous MS1 or MS5 polynucleotide sequences using a guide RNA and Cas endonuclease system to confer male sterility, allowing for precise modification of male fertility genes in plant cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chemical or random mutagenesis methods are used to induce male sterility, then male-sterile plants can be obtained, but extensive screening and backcrossing are required to eliminate undesired mutations, making the process labor-intensive and time-consuming
Solution Approach 1:
The patent replaces traditional chemical mutagenesis and random breeding methods with CRISPR/Cas9 genome editing technology. This substitution enables precise, targeted modification of male fertility genes (MS1 or MS5) without random mutations, eliminating the need for extensive screening and backcrossing. The guide RNA directs the Cas endonuclease to specifically cleave the target gene sequence, achieving reliable male sterility induction with minimal time loss.
Solution Approach 2:
The patent introduces guide RNA as an intermediary molecule that mediates between the Cas endonuclease and the target male fertility gene. The guide RNA contains a sequence complementary to the target gene, enabling specific recognition and binding. This intermediary mechanism ensures precise targeting of MS1 or MS5 genes while avoiding off-target effects, thereby achieving efficient male sterility without extensive screening.
2Productivity
If manual emasculation is used to prevent self-pollination, then hybrid seed production can be achieved, but the process becomes labor intensive and impractical for certain crops
Solution Approach 1:
The patent replaces manual emasculation operations with CRISPR/Cas9 genome editing. Instead of mechanically removing male reproductive organs, the invention uses guide RNA and Cas endonuclease to genetically disable male fertility genes (MS1 or MS5). This substitution transforms a labor-intensive mechanical process into an efficient molecular biology technique, significantly improving ease of operation while maintaining high hybrid seed production capability.
Solution Approach 2:
The patent performs preliminary genetic modification to create male-sterile plants before hybrid seed production. By using CRISPR/Cas9 to knock out MS1 or MS5 genes in advance, the female parent is pre-configured to be naturally male-sterile. This preliminary action eliminates the need for time-consuming manual emasculation during the actual hybridization process, greatly improving operational ease and productivity.
3Productivity
If traditional breeding methods are used to produce hybrid seeds, then cross-pollination can be achieved, but self-pollination of female inbred must be prevented through labor intensive procedures
Solution Approach 1:
The patent replaces complex pollination control procedures with CRISPR/Cas9 genome editing. Instead of using mechanical or chemical methods to prevent self-pollination, the invention genetically modifies the female parent to be male-sterile by disrupting MS1 or MS5 genes. This substitution simplifies the entire pollination control system, eliminating the need for manual emasculation, bagging, or other complex procedures while maintaining high hybrid seed production efficiency.
Solution Approach 2:
The patent changes the fundamental parameter of male fertility from 'fertile' to 'sterile' through CRISPR/Cas9-mediated gene disruption. By introducing specific mutations in MS1 or MS5 genes, the male fertility parameter is altered, causing the female parent to become naturally male-sterile. This parameter change simplifies pollination control procedures and dramatically improves hybrid seed production efficiency by eliminating the need for complex prevention measures.
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 male-sterile plants with reduced need for labor-intensive screening and backcrossing, ensuring targeted modification of male fertility genes, thereby improving hybrid seed production efficiency.
Implementation Method 1
providing to a plant cell a guide RNA and a Cas endonuclease. The RNA and Cas endonuclease are capable of forming a complex that enables the Cas endonuclease to introduce a double strand break at a target site located in or near a male fertility gene of MS1 or MS5
Data Source
AI summary
Compositions and methods are provided for genome modification of a nucleotide sequence located in or near a male fertility gene of Ms1 or Ms5 in the genome of a plant cell or plant to produce a male-sterile plant. In some examples, the methods and compositions employ a guide RNA/Cas endonuclease system for modifying or altering target sites located in or near a male fertility gene of Ms1 or Ms5 in the genome of a plant cell, plant or seed to produce a male-sterile plant. Also provided are compositions and methods employing a guide polynucleotide/Cas endonuclease system for genome modification a nucleotide sequence located in or near a male fertility gene of Ms1 or Ms5 in the genome of a plant cell to produce a male-sterile plant. Compositions and methods are also provided for restoring fertility to a Ms1 or Ms5 nucleotide sequence to a male-sterile Ms1 or Ms5 plant.