Nucleic Acid Sequences for Regulated Protein Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for regulating protein expression in host cells are limited by the 5'-UTR of NPT303, which can inhibit translation initiation and transcription, necessitating the development of alternative and improved strategies for controlling protein expression.
Innovation Solution
A nucleic acid construct comprising specific nucleotide sequences, including GAA repeats, TC-rich, A-rich, and GT-rich regions, along with intron sequences, is used to enhance mRNA usage and translation, allowing for regulated expression of proteins in various host cells such as yeast, fungal, and plant cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the 5'-UTR of NPT303 is used for regulating protein expression, then translation control can be achieved, but transcription is inhibited and translation efficiency is limited
Solution Approach 1:
The patent extracts the translation control function from the problematic 5'-UTR of NPT303 by identifying and isolating the specific cis-acting elements (GAA repeats, TC-rich, A-rich, and GT-rich sequences) that mediate translational regulation. These elements are then transferred to new 5'-UTR sequences that lack the transcription-inhibiting properties of the original NPT303 5'-UTR, thereby separating the useful translation control function from the harmful transcription inhibition.
Solution Approach 2:
The patent modifies the nucleotide sequences by changing specific parameters such as the composition of GAA repeats, TC-rich regions, A-rich regions, and GT-rich regions. These parameter changes create new 5'-UTR sequences that maintain translation control capability while eliminating the transcription inhibition problem. The sequences are optimized with specific repeat numbers and compositional ratios to achieve desired expression levels without harmful effects.
2Object-generated harmful factors
If alternative nucleic acid sequences are designed to overcome NPT303 limitations, then transcription inhibition is avoided, but sequence design complexity increases
Solution Approach 1:
The patent segments the translation control function into distinct modular elements: GAA repeat sequences, TC-rich sequences, A-rich sequences, and GT-rich sequences. Each segment can be independently designed and assembled into different 5'-UTR configurations. This modular segmentation simplifies the design process by allowing researchers to mix and match predefined functional modules rather than designing entirely new sequences from scratch.
Solution Approach 2:
The patent creates universal 5'-UTR sequences containing cis-acting elements that can regulate translation of various heterologous proteins across different expression systems. The designed sequences serve multiple functions: they provide translation control, avoid transcription inhibition, and can be applied to diverse protein targets. This universality reduces design complexity by establishing a reusable framework that works across multiple applications.
Data Source
AI summary
The present invention describes mRNA usage improving and/or translation-enhancing nucleic acid sequences, nucleic acid constructs comprising such sequences, and host cells comprising such nucleic acid constructs. The invention further pertains to a method for expressing a protein of interest in a cell or organism using such nucleic acid sequences, as well as their uses for increasing intergration of such nucleic acid construct into a genome, for enhancing mRNA usage and/or translation of a recombinantly expressed polypeptide, and for increasing the number of transformants upon transformation of a cell with such nucleic acid construct.