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Experimental RNAi02:15

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
RNA-seq03:21

RNA-seq

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. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
RNA Interference01:23

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
RNA Interference01:23

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
RNA Stability01:53

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
RNA Stability01:53

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...

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Video Experimental Relacionado

Updated: May 29, 2026

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
13:00

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

Published on: December 2, 2009

Optimización de nucleósidos para RNAi: una plataforma de alto rendimiento.

Gabor Butora1, Denise M Kenski, Abby J Cooper

  • 1Department of Process Chemistry, Merck & Co., Rahway, New Jersey 07065, USA. gabor_butora@merck.com

Journal of the American Chemical Society
|September 28, 2011
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores modificaron la columna vertebral del hilo guía (GS) en los complejos de silenciamiento inducidos por ARN (RISC) para mejorar la estabilidad del siRNA y el potencial terapéutico. Este enfoque sistemático permite el diseño racional de terapias de interferencia de ARN potentes, seguras y efectivas.

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Área de la Ciencia:

  • Biología Molecular Biología Molecular
  • Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN Terapéutica de interferencia de ARN
  • Descubrimiento de Drogas Descubrimiento de Drogas

Sus antecedentes:

  • El complejo de silenciamiento inducido por ARN (RISC) utiliza una hebra guía (GS) para la escisión de ARNm específica de la secuencia.
  • La modificación de la columna vertebral GS ofrece una estrategia para modular la actividad de RISC y mejorar las propiedades del siRNA.

Objetivo del estudio:

  • Evaluar sistemáticamente los nucleósidos modificados dentro de la GS de RISC.
  • Establecer una plataforma para el diseño racional de ARN pequeños interferentes mejorados (siRNA).

Principales métodos:

  • Síntesis de fosforamiditas para las cuatro bases canónicas con modificaciones.
  • Evaluación secuencial de nucleósidos modificados en cada posición de un 21-nucleótido GS.
  • Utilizó la inosina como línea de base para la comparación de nucleósidos modificados con azúcar.
  • Validación de la plataforma utilizando la modificación 2'-O-bencilo en posiciones específicas.

Principales resultados:

  • Demostró que las posiciones 5, 8, 15 y 19 de la GS pueden acomodar modificaciones voluminosas como 2'-O-benzil.
  • Estableció una metodología de alto rendimiento para evaluar nucleósidos modificados.
  • Se definió una nueva línea de base de actividad utilizando sustituciones de inosina.

Conclusiones:

  • La metodología de alto rendimiento desarrollada facilita el diseño basado en hipótesis de siRNAs.
  • Permite la creación de siRNA potentes, inmunológicamente silenciosos y estables para uso terapéutico.
  • Avanza en el desarrollo de terapias basadas en la interferencia de ARN de próxima generación.