Chimeric RNA MicroRNA Tags for Tissue-Specific Gene Expression Control
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Solution Overview
Problem
Current gene expression systems face challenges in achieving tissue-specific and temporal control, leading to leaky expression and unintended effects in applications such as plant nematode resistance and cancer therapy, due to the lack of precise spatial and temporal regulation of gene expression.
Innovation Solution
A method involving the use of chimeric RNA sequences with a nucleotide sequence linked to a microRNA tag that is complementary to naturally occurring microRNAs, allowing for enhanced specificity by modulating expression in specific tissues and conditions, thereby reducing unwanted expression in other tissues or times.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional gene expression systems are used, then gene expression can be achieved, but tissue-specific and temporal control is insufficient leading to leaky expression
Solution Approach 1:
The patent introduces microRNA tags as intermediary molecules that mediate between the transgene and the endogenous microRNA system. These tags are incorporated into the 3' UTR of the transgene transcript, allowing endogenous microRNAs to specifically regulate transgene expression in a tissue-specific and temporal manner, thereby achieving precise control without leaky expression
Solution Approach 2:
The patent changes the regulatory parameters of gene expression by introducing microRNA recognition elements (tags) that alter how the transgene transcript is regulated. By designing tags with specific complementarity to endogenous microRNAs, the system achieves dynamic control of expression levels based on spatial (tissue type) and temporal (developmental stage) parameters
2Productivity
If gene expression is enhanced for therapeutic effect, then efficacy improves, but unintended effects and toxicity increase
Solution Approach 1:
The patent applies local quality by restricting transgene expression to specific tissues and developmental stages through microRNA-mediated regulation. The microRNA tags ensure that high-level expression occurs only where and when needed, preventing systemic toxicity while maintaining high therapeutic efficacy at the target site
Solution Approach 2:
The patent employs preliminary anti-action by using microRNA tags to pre-establish regulatory control over transgene expression. The endogenous microRNA system is harnessed to suppress expression in inappropriate tissues or stages before harmful effects can occur, preventing toxicity proactively
3Productivity
If constitutive promoters are used for high expression, then productivity increases, but spatial and temporal regulation is lost
Solution Approach 1:
The patent merges constitutive expression elements with microRNA-responsive regulatory elements. The transgene is designed with a constitutive promoter for high expression potential, combined with microRNA tags in the 3' UTR that provide spatial and temporal control, achieving both high productivity and adaptability simultaneously
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 precise regulation of gene expression, minimizing unintended effects and improving the efficacy of transgenic plants and gene therapy by ensuring targeted expression, reducing toxicity and enhancing the specificity of therapeutic effects.
Implementation Method 1
a nucleotide sequence to be expressed into a chimeric RNA sequence, said nucleotide sequence comprising i) at least one sequence capable to confer a preferred phenotype or beneficial effect to said eukaryotic organism, and ii) at least one sequence substantially complementary to a microRNA sequence naturally expressed in said eukaryotic organism
Data Source
AI summary
The present invention is in the field of genetics, especially plant genetics, and provides agents capable of controlling gene expression. The present invention specifically provides sequences of naturally occurring, tissue-specifically expressed microRNAs. The invention further provides for transgenic expression constructs comprising sequences encoding such microRNAs. By incorporation of the microRNA encoding sequence the expression from the expression construct is specifically silenced in the tissue where the naturally occurring microRNA is naturally expressed. Thereby the expression profile resulting from the promoter is modulated and leakiness is reduced. The invention further provides for a method for modulating transgenic expression by incorporating sequences encoding the microRNAs into transgenic expression constructs. The compositions and methods of the invention can be used to enhance performance of agricultural relevant crops and for therapy, prophylaxis, research and diagnostics in diseases and disorders, which afflict mammalian species.