mRNA Intercellular Mobility via tRNA-Like Structures
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Solution Overview
Problem
Current understanding of mRNA transport in plants and animals is limited, particularly regarding the mechanisms and motifs that mediate long-distance intercellular mobility of mRNA transcripts, which is crucial for development, growth, and communication between cells, including potential roles in disease progression and oncogenesis.
Innovation Solution
Modifying or including tRNA-like structures (TLS) in mRNA transcripts to alter their intercellular mobility, either by mutating the gene to prevent or promote mobility, or by adding TLS sequences to change the destination of the transcripts within an organism, using genetic modification techniques such as CRISPR-Cas or transgenic methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If tRNA-like structures are added to mRNA transcripts to enhance intercellular mobility, then the ability of mRNA to move between cells is improved, but the complexity of the genetic modification process increases
Solution Approach 1:
The patent uses tRNA-like structures as intermediary elements that mediate the transport of mRNA between cells. These TLS act as molecular mediators that facilitate the movement of mRNA through cellular barriers, enabling intercellular communication without requiring complex viral vectors or transfection mechanisms.
Solution Approach 2:
The patent modifies specific parameters of the mRNA molecule by adding tRNA-like structures at defined positions (such as at the 3' end or within the coding sequence). This changes the physical and biological parameters of the mRNA, specifically its ability to interact with cellular transport mechanisms, thereby enhancing mobility without altering the core coding sequence.
2Speed
If gene mutation is used to modify tRNA-like structures in mRNA, then intercellular mobility is changed, but the precision required for genetic modification increases
Solution Approach 1:
The patent employs CRISPR-Cas9 technology to perform preliminary genetic modifications by designing guide RNAs that target specific sequences flanking the tRNA-like structure regions. This allows for precise insertion, deletion, or modification of TLS sequences before the mRNA is transcribed, ensuring the correct structural changes are made in advance.
Solution Approach 2:
The patent replaces traditional mechanical or chemical methods of genetic modification (such as restriction enzymes or chemical mutagenesis) with a molecular biology-based system using CRISPR-Cas9. This substitution enables more precise and programmable modifications of the tRNA-like structures in the mRNA gene sequences.
3Productivity
If tRNA-like structures are included in mRNA to control mobility, then protein production in specific tissues is improved, but the complexity of detecting and measuring mobility increases
Solution Approach 1:
The patent incorporates fluorescent protein tags (such as GFP or RFP) fused to the protein of interest or to the mRNA itself. This allows for visual detection and tracking of mRNA mobility and protein production in specific tissues using fluorescence microscopy, transforming the invisible molecular processes into observable color signals.
Solution Approach 2:
The patent uses reporter genes (such as luciferase or GFP) that create a measurable copy or signal of the mRNA's presence and activity. By linking the reporter gene expression to the mobility-controlled mRNA, researchers can indirectly measure mobility through the easier-to-detect reporter signal in different tissue locations.
Data Source
AI summary
The present invention relates to a method for changing the intercellular mobility of an mRNA of a gene in an organism, comprising: modifying at RNA-like structure present in the mRNA by mutating the gene from which them RNA is transcribed, or including the sequence of at RNA-like structure in the transcribed part of the gene. The method is in particular suited for plants. The intercellular mobility can be between different organs. Mutating the gene is for example for inducing loss of mobility of the transcript and comprises deleting the sequence of the tRNA-like structure from the gene, mutating the sequence of the tRNA-like structure to change the tridimensional configuration thereat; or inserting a genetic element into the gene to remove the tRNA-like structure from the transcribed part of the gene, or for inducing a change in the destination of the transcript and comprises modifying the sequence of the tRNA-like structure from the gene such that the transcript is addressed to a location different from the original destination of the transcript.