Key result
An in vitro translation system from yeast programmed by CrPV IGR IRES-containing mRNA successfully synthesized active nanoLuciferase, requiring a high eEF1A to ribosome ratio for efficient initiation.
The study developed a functional in vitro yeast translation system that operates without initiation factors by utilizing CrPV IGR IRES-containing mRNA.
Extends yeast-based tools for factor-independent translation studies; leaves open relevance to mammalian systems or clinical translation.
We developed an in vitro translation system from yeast, reconstituted with purified translation elongation and termination factors and programmed by CrPV IGR IRES-containing mRNA, which functions in the absence of initiation factors. The system is capable of synthesizing the active reporter protein, nanoLuciferase, with a molecular weight of 19 kDa. The protein synthesis by the system is appropriately regulated by controlling its composition, including translation factors, amino acids and antibiotics. We found that a high eEF1A concentration relative to the ribosome concentration is critically required for efficient IRES-mediated translation initiation, to ensure its dominance over IRES-independent random internal translation initiation.
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Abe et al. (2020) studied this question. In vitro translation system from yeast programmed by CrPV IGR IRES-containing mRNA was evaluated on Synthesis of active reporter protein, nanoLuciferase. An in vitro translation system from yeast programmed by CrPV IGR IRES-containing mRNA successfully synthesized active nanoLuciferase, requiring a high eEF1A to ribosome ratio for efficient initiation.
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