mRNA is a new medical modality as vaccine and raises hopes to become a protein replacement agent. Despite notable advances in manufacturing, stabilization and modification, its short half-life remains a major limitation. Circular RNA bears potential to solve this issue, as degradation of mRNA proceeds primarily from the 5' and 3' ends. However, circular RNAs normally lack a 5' cap and are less efficiently translated. We present a strategy to circularize mRNA covalently via click chemistry by modifying the 5' cap and the 3' poly(A) tail with bioorthogonal functional groups. The resulting Cyclic Click Cap-containing mRNA (CyCliCap-mRNA) is translationally muted by a photocleavable protecting group. Upon irradiation by light, the photocleavable protecting group is removed and the respective linear Cap0-mRNA released. We show that CyCliCap-mRNA can be activated for efficient translation at late timepoints (24-48 h post transfection) in HeLa cells, which is not possible with the respective linear mRNA. The light-mediated opening releases linear mRNA with a native Cap0 and thus overcomes the problem of inefficient translation of circular RNAs.
A new chemically regulated circRNA translation system based on small ligand-induced stabilization of RNA-binding proteins (RBPs), engineered by incorporating RNA motifs into an internal ribosomal entry site and fusing RBPs with small ligand-stabilized conditional proteins is developed.
The versatility of the assay is demonstrated and a 4-fold reduction in 5' cap concentration for in vitro transcription results in equivalent translation in cellulo, illustrating how split luminescent tagging can be easily integrated into the mRNA therapeutic workflow, enabling monitoring of real-time mRNA-driven protei...
C. Batho, Camilla Ascanelli, Megan L. Maple et al.· Journal of Visualized Experi...· 0 citations
Both acylated mRNA models resulted in significant increases in protein expression compared to the unmodified mRNA control, with increases in protein expression of up to 600% and sustained higher protein expression of modified mRNA samples observed over 120 h in comparison to unmodified RNA.
Mary E. Flood, Sathishkumar Kurusamy, Mark Berney et al.· Angewandte Chemie· 0 citations
This review article provides a comprehensive overview of current mRNA development pipelines and ongoing clinical trials, and recent advances in mRNA purification techniques, focusing on strategies for dsRNA removal.
Jin-He Liu, Jian Zang, Jing-Ru Xu et al.· Journal of Biomedical Scienc...· 0 citations
This work shows that short, chemically synthesized RNA oligonucleotides lacking some or all of these features are efficiently translated when they encode epitopes recognized by CD8+ T cells and introduces the possibility that defective or supposedly non-coding RNA may encode human and murine major histocompatibility co...
Julia Frei, D. Eisel, N. Jarzębska et al.· Nature Biomedical Engineerin...· 1 citation
A cap-independent translation enhancer BBV is introduced and protein-encoding RNA oligonucleotides (PEOs) are developed, chemically synthesized RNAs bearing 5’-OH and 3’-P termini that drive rolling circle translation after endogenous circularization that built an IVT-free platform for rapid, safe, and highly druggable...
Qian Pan, Chi Zhang, Xin-Yue Wang et al.· Proceedings of the National...· 0 citations
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