Chimeric ta-siRNA for Multi-Gene Silencing in Plants
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Solution Overview
Problem
Current methods for achieving parallel suppression or silencing of multiple genes in plants are inefficient, particularly those based on chimeric antisense molecules and double-stranded RNA molecules, which are laborious to obtain and not effective for targeting two or more genes simultaneously.
Innovation Solution
A method involving the introduction or expression of chimeric ribonucleotide sequences with modified ta-siRNA sequences, where one phase region is replaced with a sequence complementary to the target gene, and optionally the microRNA binding site is replaced with a sequence capable of mediating cleavage, such as microRNAs or siRNAs, to achieve targeted gene silencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If chimeric antisense molecules or double-stranded RNA molecules are used for gene silencing, then gene expression can be suppressed, but the methods are laborious to obtain and ineffective for targeting two or more genes simultaneously
Solution Approach 1:
The invention creates a universal chimeric RNA molecule structure that can simultaneously target multiple genes through different sequence regions. The molecule contains a conserved microRNA binding site that recruits processing enzymes, followed by variable phase regions that can be designed to match different target genes, enabling one molecule to silence multiple genes at once
Solution Approach 2:
The chimeric RNA molecule is segmented into functional domains: a microRNA binding site region for enzyme recruitment, phase regions for gene-specific targeting, and structural elements for stability. This segmentation allows independent optimization of each function and facilitates targeting of multiple genes by varying the phase region sequences
2Productivity
If current methods are used for parallel suppression of multiple genes, then some gene silencing can be achieved, but the efficiency is low and the process is not effective for simultaneous targeting
Solution Approach 1:
The invention merges the gene-silencing function of siRNA with the amplification capability of microRNA pathways. By designing chimeric molecules that contain both microRNA binding sites and gene-specific phase regions, the system combines the specificity of siRNA with the efficiency of microRNA-mediated RNAi, achieving high-efficiency simultaneous silencing of multiple genes
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 allows for efficient and specific silencing of multiple target genes in plants, including those involved in developmental processes and pathogen regulation, offering improved agronomic traits like disease resistance and stress tolerance.
Implementation Method 1
one phase region is replaced by a sequence, which is substantially complementary to said target gene
Data Source
AI summary
The present invention is in the field of genetics, especially plant genetics, and provides agents capable of controlling gene expression. More specifically the inventions relates to methods for engineering ta-siRNA primary transcripts in order to target gene-of-interest (GOI) and control their expression. The invention further provides for a method for modulating transgenic expression by said engineered ta-siRNAs.