Design of an affordable RNA system that starts protein production without needing usual start signals

The novel RNA construct achieved comparable expression efficiency and stability to conventional mRNAs without requiring a 5' cap or a 3' poly(A) tail.

• In vitro mRNA transcription typically requires a 5' cap and a 3' poly(A) tail for protein expression in eukaryotic cells.
• Replacing the 5' cap and untranslated region with an internal ribosome entry site (IRES) from the encephalomyocarditis virus (EMCV) may simplify production.
• A replication-dependent histone stem-loop structure was used to substitute the 3' poly(A) tail.
• The β-globin 3' UTR combined with the Homo sapiens histone SL was identified as the optimal configuration for efficient translation.
• The new mRNA construct demonstrated expression efficiency and stability comparable to traditional capped and polyadenylated mRNAs (p > 0.05).
• This approach may reduce complexity and cost in industrial-scale mRNA production.

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