Rice Flotillin1 and Importin α4 Gene Knockout for RSV Resistance
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Solution Overview
Problem
Plant virus infections, particularly those caused by rice stripe virus (RSV), pose significant economic threats to rice yields, with existing research focusing on protein interactions and host immunity but lacking clear understanding of viral receptors, necessitating a method to inhibit virus spread and enhance plant resistance.
Innovation Solution
Knockout of the Flotillin1 and Importin α4 genes in rice using CRISPR/Cas9 technology to reduce their expression, employing sgRNAs to target and edit these genes, thereby inhibiting protein production and enhancing resistance to RSV.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If Flotillin1 and Importin α4 genes are knocked out to enhance virus resistance, then the anti-disease capability is improved, but the complexity of genetic modification increases
Solution Approach 1:
The patent segments the virus resistance enhancement into two distinct gene targets (Flotillin1 and Importin α4), allowing researchers to address the complex problem of virus resistance through multiple manageable genetic modifications rather than a single complex intervention
Solution Approach 2:
The patent uses CRISPR/Cas9 technology as an intermediary tool to achieve gene knockout, replacing more complex traditional genetic modification methods with a streamlined molecular mechanism that guides precise gene editing through RNA-mediated targeting
2Object-affected harmful factors
If gene knockout is performed to reduce virus spread, then the disease incidence is reduced, but the difficulty of detecting and measuring gene function increases
Solution Approach 1:
The patent incorporates feedback mechanisms by using quantitative PCR and western blotting to monitor gene expression levels and protein abundance, allowing researchers to verify the success of gene knockout and assess its impact on virus resistance through measurable molecular markers
Solution Approach 2:
The patent replaces traditional mechanical or visual observation methods for assessing gene function with molecular biology techniques (PCR, western blotting) that provide quantitative and objective measurement of gene knockout efficiency and its biological consequences
3Productivity
If Flotillin1 and Importin α4 expression is reduced to inhibit RSV spread, then the virus replication is suppressed, but the loss of substance (gene function) increases
Solution Approach 1:
The patent extracts and removes the specific gene sequences (Flotillin1 and Importin α4) that facilitate virus replication, using CRISPR/Cas9 to precisely excise or disable these genetic elements while leaving the rest of the genome intact, thereby suppressing virus replication without causing widespread genetic disruption
Solution Approach 2:
The patent applies local quality modification by targeting specific genes (Flotillin1 and Importin α4) with distinct molecular functions that are critical for virus replication, rather than applying blanket suppression to all genes, thereby achieving virus replication suppression while minimizing loss of overall genetic function
Data Source
AI summary
The present application relates to the field of biotechnology, in particular to the use of Flotillin1 gene and Importin α4 gene and the use of molecules for inhibiting the transcription or translation of Flotillin1 gene and Importin α4 gene in the manufacture of kits. The present application also relates to a method for obtaining plants resistant to pathogens.