mRNA Histone Stem-Loop Poly(A) Hybrid Enhances Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current gene therapy and genetic vaccination methods face challenges in achieving sufficient protein expression due to the instability of RNA molecules and limited translational efficiency, which can impact the effectiveness of these therapeutic approaches.
Innovation Solution
Incorporating a nucleic acid sequence that combines a coding region with a histone stem-loop and a poly(A) sequence or polyadenylation signal, which synergistically enhances protein expression by stabilizing the mRNA and increasing translational efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional mRNA sequences are used for gene therapy, then the therapeutic approach can be implemented, but protein expression levels are insufficient due to RNA instability and limited translational efficiency
Solution Approach 1:
The patent combines two different 3' end structures - the histone stem-loop (typically associated with replication-dependent histone genes) and the poly(A) sequence (typical of most eukaryotic mRNAs) - into a single hybrid structure. This merging creates a chimeric 3' end that leverages the stability properties of the poly(A) tail while incorporating the translational enhancement features of the stem-loop structure, thereby simultaneously improving both mRNA stability and protein expression levels
Solution Approach 2:
The invention creates a composite RNA structure by integrating sequences from different functional categories. The 3' end comprises both the poly(A) sequence (providing stability through protection from exonucleases and promotion of translation) and the stem-loop structure (providing additional stability and translational enhancement). This composite structure functions as a unified element that delivers synergistic benefits greater than the sum of its individual components
2Productivity
If conventional gene therapy methods are used, then treatment can be administered, but translational efficiency is limited reducing therapeutic effectiveness
Solution Approach 1:
The patent applies local quality modification by specifically enhancing the 3' end region of the mRNA molecule with the hybrid stem-loop/poly(A) structure. Rather than modifying the entire mRNA sequence, the invention focuses translational enhancement elements at the critical 3' terminus where they can most effectively interact with translation initiation factors and ribosomes, thereby improving translational efficiency without unnecessarily complicating the overall nucleic acid construct
Data Source
AI summary
The present application describes a coding nucleic acid sequence, particularly a messenger RNA (mRNA), comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal and the use thereof for increasing the expression of an encoded protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine e.g. for the use in the treatment of tumours and cancer diseases, cardiovascular diseases, infectious diseases, autoimmune diseases or genetic diseases, or in gene therapy. The present invention further describes an in vitro transcription method, in vitro methods for increasing the expression of a protein using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal and an ex vivo and in vivo method.