Histone Stem-Loop and Poly(A) Sequence for mRNA Expression
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Solution Overview
Problem
Current gene therapy and genetic vaccination methods face challenges in achieving sufficient expression levels of encoded proteins, particularly due to the instability and limited translational efficiency of RNA-based applications compared to DNA vaccines, which affects their therapeutic efficacy in treating allergies and autoimmune diseases.
Innovation Solution
A nucleic acid sequence comprising a coding region for a peptide or protein with an allergenic antigen or autoimmune self-antigen, combined with a histone stem-loop and a poly(A) sequence or polyadenylation signal, enhances protein expression by synergistic action, improving mRNA stability and translational efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If RNA-based gene therapy or genetic vaccination is used, then therapeutic treatment for allergies and autoimmune diseases is enabled, but mRNA stability and translational efficiency are insufficient
Solution Approach 1:
The patent combines two different 3' end structures - the histone stem-loop structure and the poly(A) sequence - into a single mRNA molecule. This merging of structures synergistically improves mRNA stability and translational efficiency, resolving the contradiction between enabling RNA-based therapy and achieving sufficient stability. The combined structure leverages the advantages of both the stem-loop (protection from exonucleases) and poly(A) sequence (enhanced translation and stability).
Solution Approach 2:
The invention creates a composite mRNA structure by integrating the histone stem-loop element and poly(A) tail into a unified 3' end architecture. This composite structure combines the protective function of the stem-loop with the stabilizing and translational enhancement functions of the poly(A) sequence, achieving superior therapeutic efficacy compared to either structure alone.
2Reliability
If RNA-based gene therapy is used, then treatment for allergies and autoimmune diseases is enabled, but translational efficiency is limited
Solution Approach 1:
The patent merges the histone stem-loop structure with the poly(A) sequence at the 3' end of the mRNA, creating a hybrid structure that simultaneously enhances translational efficiency and protein expression. The poly(A) sequence recruits poly(A)-binding proteins that facilitate ribosome recruitment and translation initiation, while the stem-loop provides structural stability, together resolving the limitation of translational efficiency in RNA-based therapies.
Solution Approach 2:
The invention modifies the 3' end structure parameter of the mRNA by introducing the combined stem-loop and poly(A) configuration. This structural parameter change fundamentally alters the mRNA's interaction with cellular translation machinery, significantly improving translational efficiency and protein expression levels compared to conventional mRNA designs.
3Ease of manufacture
If conventional mRNA structure is used, then genetic vaccination is simplified, but protein expression levels are insufficient
Solution Approach 1:
The patent combines the histone stem-loop and poly(A) sequence into the mRNA construct, achieving high protein expression levels without complicating the manufacturing process. The modular nature of this combined structure allows it to be easily incorporated into existing vaccine production platforms, maintaining ease of manufacture while dramatically improving protein expression yields.
Solution Approach 2:
The invention optimizes the 3' end structural parameter of the mRNA by implementing the stem-loop-poly(A) hybrid structure. This parameter optimization increases protein expression levels by enhancing mRNA stability and translational efficiency, while the structure remains compatible with standard genetic vaccination manufacturing workflows.
Data Source
AI summary
The present invention relates to a nucleic acid sequence, comprising or coding for a coding region, encoding at least one peptide or protein comprising an allergenic antigen or an autoimmune self-antigen or a fragment, variant or derivative thereof, at least one histone stem-loop and a poly(A) sequence or a polyadenylation signal. Furthermore the present invention provides the use of the nucleic acid for increasing the expression of said encoded peptide or protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine, e.g. for use in the treatment of allergies or autoimmune diseases. The present invention further describes a method for increasing the expression of a peptide or protein comprising an allergenic antigen or an autoimmune self-antigen or a fragment, variant or derivative thereof, using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal.