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Related Concept Videos

Leaky Scanning02:28

Leaky Scanning

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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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Initiation of Translation02:33

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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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Ribosomal RNA Synthesis02:53

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Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
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Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
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Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

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RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
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Related Experiment Video

Updated: Jun 5, 2025

Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli
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Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli

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Efficient circular RNA synthesis for potent rolling circle translation.

Yifei Du1, Philipp Konrad Zuber2, Huajuan Xiao3

  • 1MRC Laboratory of Molecular Biology, Cambridge, UK. yifei.du18@gmail.com.

Nature Biomedical Engineering
|December 13, 2024
PubMed
Summary
This summary is machine-generated.

New trans-splicing methods enable the synthesis of large, stable circular RNAs (circRNAs) for therapeutic applications. These novel circRNA constructs show enhanced translation and low immunogenicity, paving the way for advanced RNA therapeutics.

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Area of Science:

  • Molecular Biology
  • RNA Therapeutics

Background:

  • Circular RNA (circRNA) is a promising candidate for next-generation RNA therapeutics due to its inherent stability.
  • Existing methods for circRNA synthesis have limitations in scale, efficiency, and adaptability.

Purpose of the Study:

  • To develop novel trans-splicing-based methods for synthesizing large circRNAs (>8,000 nucleotides).
  • To evaluate the therapeutic potential of these synthesized circRNAs, focusing on immunogenicity and translation efficiency.

Main Methods:

  • Employed trans-splicing strategies for circRNA synthesis, independent of bacterial sequences.
  • Incorporated RNA modifications and human 28S ribosomal RNA sequences.
  • Utilized viral internal ribosomal entry sites for rolling circle translation.

Main Results:

  • Successfully synthesized large circRNAs (>8,000 nt) with high efficiency and reliability.
  • Unmodified circRNAs demonstrated low immunogenicity and superior translation efficiency compared to those from permuted intron-exon methods.
  • Achieved over 7,000-fold enhancement in rolling circle translation efficiency.

Conclusions:

  • The developed trans-splicing methods offer a robust platform for producing therapeutic-grade circRNAs.
  • These findings support the advancement of circRNA technology for potential clinical applications in RNA therapeutics.