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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
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Regulation of Expression at Multiple Steps01:23

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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
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One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Translational Regulation

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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Riboswitches01:56

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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Related Experiment Video

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Quantitative Immunofluorescence to Measure Global Localized Translation
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Synthesis of Precisely Modified Ribonucleic Acid to Reveal the Site-Specific Effect of Modification on Message

Yufan Pan1, Chenyou Zhu1, Yuan Zhuang1,2

  • 1Engineering Research Center of Advanced Rare Earth Materials (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing, 100084, China.

Journal of the American Chemical Society
|October 16, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method for creating site-specifically modified long single-strand RNA (ssRNA). This advancement allows precise control over RNA modifications, enhancing mRNA therapy safety and efficacy.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Development

Background:

  • RNA modifications are crucial for mRNA therapy development.
  • Concerns exist regarding site-specific modification accuracy and biosafety.
  • Understanding precise modification effects is vital for advancing mRNA therapeutics.

Purpose of the Study:

  • To develop a template-directed strategy for synthesizing site-specifically modified long ssRNA.
  • To investigate the impact of precisely modified mRNA patterns on translation.
  • To provide a guideline for mRNA modification in next-generation therapeutics and RNA epigenetics.

Main Methods:

  • Developed a template-directed synthesis strategy for long ssRNA (over 300 nt).
  • Achieved single-base resolution modifications including m6A, m5C, m1Ψ, Ψ, I, Br-dU, Cy3, Cy5, FAM, 2'-F, 2'-OMe, 2'-MOE, 2'-Propargyl, LNA, cET, and PS.
  • Investigated the functional impact of specific modification patterns on mRNA translation.

Main Results:

  • Successfully synthesized long ssRNAs with site-specific modifications at single-base resolution.
  • Demonstrated that m1Ψ modification at specific sites enhances translation fidelity.
  • Observed that m1Ψ modification retains low immunogenicity while improving translation.

Conclusions:

  • The developed strategy enables precise control over RNA modifications for long ssRNA.
  • Site-specific m1Ψ modification offers a promising approach to improve mRNA translation fidelity and safety.
  • This research provides a foundation for developing advanced mRNA drugs and advancing RNA epigenetics.