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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
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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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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Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
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Updated: Jun 15, 2025

De novo Identification of Actively Translated Open Reading Frames with Ribosome Profiling Data
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Descubrimiento de ORFs panvirales utilizando perfiles de ribosomas masivamente paralelos

Shira Weingarten-Gabbay1,2,3, Matthew R Bauer4, Alexandra C Stanton1,5,6

  • 1Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Science (New York, N.Y.)
|June 12, 2025
PubMed
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Los científicos descubrieron miles de nuevos marcos de lectura abiertos virales (ORF) utilizando perfiles masivos de ribosomas paralelos (MPRP). Estos hallazgos revelan nuevos péptidos virales y elementos reguladores, expandiendo los objetivos de la vacuna y entendiendo los mecanismos virales.

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Área de la Ciencia:

  • Virología
  • La genómica
  • Inmunología

Sus antecedentes:

  • La definición de los proteomas virales es esencial para comprender los ciclos de vida virales y las respuestas inmunes.
  • La extensión total de las regiones traducidas en los genomas virales es en gran medida desconocida.

Objetivo del estudio:

  • Desarrollar y aplicar un método para la identificación de nuevos marcos de lectura abierta de virus (ORF).
  • Caracterizar el paisaje de las regiones traducidas en genomas virales asociados a humanos.

Principales métodos:

  • Se empleó el perfil masivo de ribosomas paralelos (MPRP) para analizar decenas de miles de oligonucleótidos diseñados.
  • El MPRP se utilizó para identificar y mapear los ORF en numerosos genomas virales.

Principales resultados:

  • Se identificaron 4208 ORF no anotados en 679 genomas virales asociados con humanos.
  • Los péptidos virales de los ORF no canónicos se presentaron en moléculas de clase I del antígeno leucocitario humano (HLA).
  • Se encontraron cientos de ORFs upstream (uORFs), que podrían regular la traducción de proteínas virales.

Conclusiones:

  • El descubrimiento de numerosos ORF virales expande el proteoma viral conocido y los objetivos potenciales de la vacuna.
  • Los ORF y uORF identificados ofrecen información sobre las secuencias cis-reguladoras virales y la modulación de la traducción.
  • Este trabajo mejora nuestra comprensión de la biología viral y el reconocimiento inmune en diversas familias virales.