Simultaneous RNA Production via Merged In Vitro Transcription
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Solution Overview
Problem
Current RNA production methods are inefficient and costly as they require separate production processes for each specific RNA molecule species, making it laborious and expensive to produce a mixture of RNA species simultaneously, which is desirable for treatments involving multiple antigens or pathogens.
Innovation Solution
A method for producing a ribonucleic acid (RNA) molecule composition comprising multiple RNA species by in vitro transcription of a mixture of DNA molecule species in a single reaction vessel, allowing for simultaneous generation of multiple RNA species from different DNA templates, using techniques such as bacterial amplification, PCR, chemical DNA synthesis, or enzymatic amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If separate production processes are used for each RNA molecule species, then each RNA can be produced with high purity and specificity, but the production time, labor, and cost increase significantly
Solution Approach 1:
The patent combines multiple separate RNA production processes into a single in vitro transcription reaction. Multiple DNA templates (plasmids) encoding different RNA species are transcribed simultaneously in one reaction vessel, producing a mixture of RNA molecules. This merging approach maintains the purity and specificity of individual RNA species while dramatically improving production efficiency by eliminating the need for separate production processes for each RNA.
Solution Approach 2:
The in vitro transcription system is designed to handle multiple DNA templates simultaneously, making the system universal for producing various RNA species in a single reaction. The reaction system can accommodate different DNA templates (e.g., plasmids encoding viral proteins, cytokines, or other therapeutic proteins) and produce their corresponding RNA molecules in one go, thereby improving productivity without sacrificing manufacturing precision.
2Adaptability or versatility
If multiple RNA species are produced in separate processes, then each RNA can be optimized independently, but the overall production time and cost increase
Solution Approach 1:
The patent merges multiple RNA production processes into a single simultaneous in vitro transcription reaction. Multiple DNA templates can be included in one reaction vessel, allowing different RNA species to be produced at the same time. This reduces production time significantly compared to sequential production, while still allowing independent optimization of each RNA species through careful selection and design of the DNA templates.
Solution Approach 2:
The DNA templates (plasmids) are prepared and optimized in advance before the transcription reaction. This preliminary action allows the RNA species to be produced simultaneously without requiring sequential optimization during the production process. The DNA templates can be pre-designed with appropriate promoters, coding sequences, and regulatory elements, enabling efficient simultaneous transcription of multiple RNA species.
3Stability of the object's composition
If DNA is used for gene therapy, then the DNA is stable and easy to handle, but the risk of undesired genomic integration and anti-DNA antibody generation increases
Solution Approach 1:
The patent extracts the genetic information from stable DNA and converts it into RNA molecules for therapeutic application. By using RNA instead of DNA, the harmful effects of genomic integration and anti-DNA antibody generation are eliminated, while the genetic information remains stable and easy to handle through the DNA templates used in the in vitro transcription system. The DNA templates serve as stable storage for the genetic code, which is then transcribed into functional RNA molecules that do not integrate into the genome.
4Object-affected harmful factors
If mRNA is used for gene therapy, then the risk of genomic integration and anti-DNA antibody generation is minimized, but the mRNA stability was considered insufficient
Solution Approach 1:
The patent employs preliminary action by using stable DNA templates (plasmids) that are prepared in advance and optimized for transcription. These DNA templates contain all the necessary genetic information and regulatory elements, allowing for the production of stable and functional RNA molecules. The DNA templates serve as stable storage media that can be handled easily, while the resulting RNA molecules benefit from the stability provided by their DNA origins without requiring genomic integration.
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 reduces production time, labor, and costs by enabling the simultaneous production of multiple RNA species in a single batch, which is advantageous for pandemic scenarios and personalized medicine, while minimizing the risk of genomic integration and antibody generation.
Implementation Method 1
RNA in vitro transcription of a mixture of m different deoxyribonucleic acid (DNA) molecule species in a single reaction vessel
Data Source
AI summary
The invention relates to a method for producing a ribonucleic acid (RNA) molecule composition comprising n different RNA molecule species, the method comprising a step of RNA in vitro transcription of a mixture of m different deoxyribonucleic acid (DNA) molecule species in a single reaction vessel in parallel, i.e. simultaneously, and a step of obtaining the RNA molecule composition. Also provided is the RNA composition provided by the inventive method and a pharmaceutical composition comprising the same as well as a pharmaceutical container. Moreover, the invention provides the RNA composition and the pharmaceutical composition for use as medicament.