Chimeric Post-Transcriptional Regulatory Element for mRNA Expression
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Solution Overview
Problem
Current methods for maximizing protein expression, such as therapeutic antibodies and growth factors, face challenges in achieving stable and efficient mRNA expression due to limitations in post-transcriptional regulatory elements (PRE) sequences, which affect cytoplasmic accumulation and stability of mRNA.
Innovation Solution
The development of chimeric PRE sequences combining subelements from different PRE sequences, such as WPRE, HPRE, GSPRE, BPRE, and ASPRE, to create constructs with improved stability and expression efficiency by optimizing the arrangement and orientation of alpha, beta, and gamma subelements within a polynucleotide construct.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If native PRE sequences (WPRE, HPRE, GSPRE, BPRE, ASPRE) are used to regulate mRNA expression, then post-transcriptional regulation is achieved, but expression stability and efficiency are limited
Solution Approach 1:
The patent applies composite materials by creating chimeric PRE sequences that combine functional subelements from multiple native PRE sequences (WPRE, HPRE, GSPRE, BPRE, ASPRE). This composite approach integrates the stabilizing subelements from different viral PRE sequences to form a hybrid regulatory element with superior mRNA stability and expression efficiency compared to any single native PRE sequence.
Solution Approach 2:
The patent applies segmentation by dividing native PRE sequences into functional subelements (alpha, beta, gamma subelements) and recombining them in optimized arrangements. This segmentation allows identification and utilization of specific subelements responsible for stability and expression efficiency, enabling the construction of chimeric PRE sequences with tailored functions.
2Reliability
If PRE sequences are optimized for cytoplasmic accumulation, then mRNA stability improves, but expression efficiency may be compromised
Solution Approach 1:
The patent applies local quality by assigning different subelements to specific functional roles within the chimeric PRE sequence. Stabilizing subelements are positioned to maximize mRNA half-life, while other subelements are arranged to optimize transcription and translation efficiency. This localized functional optimization ensures that different regions of the PRE sequence contribute to different aspects of gene expression.
Solution Approach 2:
The patent applies parameter changes by modifying the composition and arrangement of subelements in PRE sequences. By varying the types, numbers, and positions of alpha, beta, and gamma subelements from different viral sources, the patent optimizes parameters such as mRNA stability, nuclear export efficiency, and cytoplasmic accumulation to achieve superior overall expression.
Data Source
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AI summary
The present disclosure relates to chimeric post-transcriptional regulatory elements (PRE) and vectors useful for expressing a protein in a cell. The PRE contains alpha, beta and optionally gamma subelements selected from different native PRE sequences and are discovered to be more potent than their native counterparts.