Recombinant mRNA UTR Design for Endothelial Cell Expression
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Solution Overview
Problem
Existing recombinant nucleic acid constructs, particularly mRNA, face challenges in achieving high expression levels and efficient expression in endothelial cells, limiting their therapeutic potential.
Innovation Solution
A recombinant nucleic acid construct with specific 5' and 3' untranslated regions (UTRs) from genes like MCP-1, RPL12s.c., Ang-2, HSP70, and others, along with a poly-A tail and optional elements like a cap structure or IRES sequence, to enhance expression and cellular function restoration in endothelial cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional mRNA constructs are used, then the construct can be administered, but expression levels are low and expression in endothelial cells is inefficient
Solution Approach 1:
The patent applies parameter changes by selecting specific 5' UTR and 3' UTR sequences from genes known to be highly expressed in endothelial cells (such as vWF, MCP-1, RPL12s.c., Ang-2, HSP70, Galectin-9, GADD34, EDN1, HSP70m5, E-selectin, ICAM-1, IL-6). By changing the UTR parameters to match endothelial cell-preferred sequences, the construct achieves high expression levels and efficient expression specifically in endothelial cells, resolving the contradiction between general administrability and targeted expression efficiency.
2Reliability
If the construct uses standard mRNA structure, then it maintains basic functionality, but it cannot achieve high expression or effective restoration of cellular functions in endothelial cells
Solution Approach 1:
The patent applies local quality by making the 5' UTR and 3' UTR regions specific to endothelial cell preferences while keeping the coding sequence general. The 5' UTR is selected from specific sequences (e.g., vWF 5' UTR, MCP-1 5' UTR) that are optimized for endothelial cell translation initiation, and the 3' UTR is selected from sequences (e.g., vWF 3' UTR, MCP-1 3' UTR) that enhance stability and translation in these cells. This localized optimization of UTR regions enables both high expression levels and effective restoration of cellular functions in endothelial cells.
3Ease of manufacture
If conventional UTR sequences are used, then the construct is simple to design, but expression in endothelial cells is not achieved
Solution Approach 1:
The patent maintains ease of manufacture by providing a clear selection criteria for UTR sequences from a defined set of genes (vWF, MCP-1, RPL12s.c., Ang-2, HSP70, Galectin-9, GADD34, EDN1, HSP70m5, E-selectin, ICAM-1, IL-6). The methodology is straightforward: select a 5' UTR and 3' UTR from this defined set, combine with any coding sequence, and the construct will achieve high expression in endothelial cells. This systematic approach simplifies the design process while ensuring reliable endothelial cell expression.
Data Source
AI summary
The present invention is related to a recombinant nucleic acid construct comprising in 5′→3′ directiona 5′ UTR,a coding region coding for an effector molecule, anda 3′ UTR,wherein 5′ UTR is selected from the group comprising a 5′ UTR of a gene or a derivative thereof having a nucleotide identity of at least 85%, wherein the gene is selected from the group consisting of MCP-1, RPL12s.c., Ang-2, HSP70, H3.3., Galectin-9, GADD34, EDN1, HSP70m5, E-selectin, ICAM-1, IL-6 and vWF.