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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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Transfer RNA Synthesis02:36

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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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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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Quantitative Immunofluorescence to Measure Global Localized Translation
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Síntesis de ácido ribonucleico modificado con precisión para revelar el efecto específico del sitio de la

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
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Este estudio introduce un nuevo método para crear ARN de cadena única larga modificado específicamente para el sitio (SSRNA). Este avance permite un control preciso sobre las modificaciones del ARN, mejorando la seguridad y eficacia de la terapia de ARNm.

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Área de la Ciencia:

  • La bioquímica
  • Biología molecular
  • Desarrollo de drogas

Sus antecedentes:

  • Las modificaciones de ARN son cruciales para el desarrollo de la terapia de ARNm.
  • Existen preocupaciones con respecto a la precisión y la bioseguridad de las modificaciones específicas del sitio.
  • Comprender los efectos precisos de la modificación es vital para el avance de las terapias de ARNm.

Objetivo del estudio:

  • Desarrollar una estrategia dirigida a la plantilla para la síntesis de ssRNA largo modificado específicamente para el sitio.
  • Investigar el impacto de los patrones de ARNm modificados con precisión en la traducción.
  • Proporcionar una guía para la modificación de ARNm en terapias de próxima generación y epigenética de ARN.

Principales métodos:

  • Desarrolló una estrategia de síntesis dirigida por plantilla para ssRNA largo (más de 300 nt).
  • Las modificaciones de resolución de una sola base alcanzadas incluyen m6A, m5C, m1Ψ, Ψ, I, Br-dU, Cy3, Cy5, FAM, 2'-F, 2'-OMe, 2'-MOE, 2'-Propargyl, LNA, cET y PS.
  • Se investigó el impacto funcional de los patrones de modificación específicos en la traducción del ARNm.

Principales resultados:

  • Se han sintetizado con éxito largos ssRNAs con modificaciones específicas del sitio a resolución de una sola base.
  • Demostró que la modificación m1Ψ en sitios específicos mejora la fidelidad de la traducción.
  • Se observó que la modificación m1Ψ conserva una baja inmunogenicidad al tiempo que mejora la traducción.

Conclusiones:

  • La estrategia desarrollada permite un control preciso de las modificaciones del ARN para el ARNss largo.
  • La modificación m1Ψ específica del sitio ofrece un enfoque prometedor para mejorar la fidelidad y la seguridad de la traducción de ARNm.
  • Esta investigación proporciona una base para el desarrollo de medicamentos avanzados de ARNm y el avance de la epigenética de ARN.