Design principles and applications of synthetic self‐replicating <scp>RNAs</scp>

Wagner, Alexander; Mutschler, Hannes

doi:10.1002/wrna.1803

Cited by 4 publications

(5 citation statements)

References 126 publications

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“…Supposedly, self-amplifying RNA vaccines may offer important advantages over conventional mRNA vaccines [63,64]. First, since saRNA vaccines contain a built-in RNA polymerase, their dose can be significantly reduced.…”

Section: Self-amplifying Rna Vaccines (Sarna)mentioning

confidence: 99%

Mucosal mRNA Vaccines. Merits and Perspectives

Toniolo,

Maccari,

Camussi

2024

Preprint

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This review highlights the potential and limitations of mRNA vaccines with a particular focus on mucosal delivery as an alternative to parenteral administration. The strengths of mRNA vaccines are: a) Versatility: they can be adapted to target a wide range of pathogens; b) Faster development: designed and produced more quickly than traditional vaccines; c) High efficacy: effective in preventing severe disease, as demonstrated during the COVID-19 pandemic. Current challenges: a) Delivery: current methods (mainly intramuscular injection) may not be optimal for inducing mucosal immunity; b) Stability: mRNA is fragile and needs to be protected from degradation; c) Safety: potential side effects, including myocarditis, blood clotting, allergic reactions need further investigation. Mucosal delivery could be more effective in preventing the transmission of pathogens and is generally better accepted by the population. Pros: offers the potential to induce local immunity at the site of pathogen entry, potentially providing broader protection; Cons: developing effective delivery systems and adjuvants for mucosal delivery is difficult. Extracellular vesicles as a mucosal delivery system are promising and offer a potentially safe, scalable, and needle-free approach. Future directions: a) Optimizing mRNA constructs: strategies like codon optimization and self-amplifying RNA can improve stability and translation efficiency; b) Rigorous pre-clinical studies are essential before human trials.

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Section: Self-amplifying Rna Vaccines (Sarna)mentioning

confidence: 99%

Mucosal mRNA Vaccines. Merits and Perspectives

Toniolo,

Maccari,

Camussi

2024

Preprint

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“…6.4. Self-Amplifying RNA Vaccines (saRNA) Supposedly, self-amplifying RNA vaccines may offer important advantages over conventional mRNA vaccines [112,113]. First, since saRNA vaccines contain a built-in RNA polymerase, their dose can be significantly reduced.…”

Section: Codon Usagementioning

confidence: 99%

mRNA Technology and Mucosal Immunization

Toniolo,

Maccari,

Camussi

2024

Vaccines

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Current mRNA vaccines are mainly administered via intramuscular injection, which induces good systemic immunity but limited mucosal immunity. Achieving mucosal immunity through mRNA vaccination could diminish pathogen replication at the entry site and reduce interhuman transmission. However, delivering mRNA vaccines to mucosae faces challenges like mRNA degradation, poor entry into cells, and reactogenicity. Encapsulating mRNA in extracellular vesicles may protect the mRNA and reduce reactogenicity, making mucosal mRNA vaccines possible. Plant-derived extracellular vesicles from edible fruits have been investigated as mRNA carriers. Studies in animals show that mRNA vehiculated in orange-derived extracellular vesicles can elicit both systemic and mucosal immune responses when administered by the oral, nasal, or intramuscular routes. Once lyophilized, these products show remarkable stability. The optimization of mRNA to improve translation efficiency, immunogenicity, reactogenicity, and stability can be obtained through adjustments of the 5′cap region, poly-A tail, codons selection, and the use of nucleoside analogues. Recent studies have also proposed self-amplifying RNA vaccines containing an RNA polymerase as well as circular mRNA constructs. Data from parenterally primed animals demonstrate the efficacy of nasal immunization with non-adjuvanted protein, and studies in humans indicate that the combination of a parenteral vaccine with the natural exposure of mucosae to the same antigen provides protection and reduces transmission. Hence, mucosal mRNA vaccination would be beneficial at least in organisms pre-treated with parenteral vaccines. This practice could have wide applications for the treatment of infectious diseases.

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“…However, considering their rapid evolutionary adaptation in cell-free transcription-translation systems, phage-derived srRNAs might also be effective for the directed evolution of additional gene cargo. 10 Notably, Qβ-based srRNAs are known for their susceptibility to "RNA parasites", which are spontaneously emerging, rapidly replicating short RNA molecules. 11,12 Previously, we demonstrated that the in vitro RNA replication catalyzed by the RdRP of Emesvirus zinderi (MS2) is less susceptible to the emergence of untemplated RNA parasites.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Consequently, phage-derived srRNA systems find their most significant application in in vitro evolution experiments involving RNA scaffolds and the associated RNA-dependent RNA polymerases (RdRPs). However, considering their rapid evolutionary adaptation in cell-free transcription-translation systems, phage-derived srRNAs might also be effective for the directed evolution of additional gene cargo . Notably, Qβ-based srRNAs are known for their susceptibility to “RNA parasites”, which are spontaneously emerging, rapidly replicating short RNA molecules. , …”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Design of Novel Synthetic RNA Replicons Based on Emesvirus zinderi

Wagner,

Mutschler

2024

ACS Synth. Biol.

Self Cite

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Self-replicating RNAs (srRNAs) are synthetic molecules designed to mimic the self-replicating ability of viral RNAs. srRNAs hold significant promise for a range of applications, including enhancing protein expression, reprogramming cells into pluripotent stem cells, and creating cell-free systems for experimental evolution. However, the development of srRNAs for use in bacterial systems remains limited. Here, we demonstrate how a srRNA scaffold from Emesvirus zinderi can be engineered into a self-encoding srRNA by incorporating the coding region of the catalytically active replicase subunit. With the help of in vitro replication assays, including an in vitro translation-coupled replication approach, we show that the resulting system enables complete replication cycles of RNA both in cis and trans, including long cargo RNAs such as tethered 5S, 16S, and 23S rRNAs. In summary, our findings suggest that these srRNAs have significant potential for fundamental research, synthetic biology, and general in vitro evolution.

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Design principles and applications of synthetic self‐replicating RNAs

Cited by 4 publications

References 126 publications

Mucosal mRNA Vaccines. Merits and Perspectives

Mucosal mRNA Vaccines. Merits and Perspectives

mRNA Technology and Mucosal Immunization

Design of Novel Synthetic RNA Replicons Based on Emesvirus zinderi

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