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RNA Structure01:23

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The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
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The basic structure of RNA consists of a string of ribonucleotides attached by phosphodiester bonds. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
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There are three main types of ribonucleic acid (RNA) involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three...
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Nucleic Acid Structure01:25

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The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms  a 5′ to 3′ phosphodiester linkage.
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in regulating gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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RNA Editing02:23

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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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DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
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Funcionalización de ARN selectivo por el sitio a través de la estructura inducida por ADN

Lu Xiao1, Maryam Habibian1, Eric T Kool1

  • 1Department of Chemistry, ChEM-H Institute and Stanford Cancer Institute, Stanford University, Stanford, California 94305, United States.

Journal of the American Chemical Society
|September 1, 2020
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores desarrollaron la acilación del ARN en bucles inducidos (RAIL), un método dirigido por el ADN para la funcionalización del ARN específico del sitio. Esta técnica permite etiquetar y controlar con precisión las moléculas de ARN, superando las limitaciones de los métodos estocásticos.

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Área de la Ciencia:

  • La bioquímica
  • Biología molecular
  • Biología Química

Sus antecedentes:

  • La funcionalización del ARN específico del sitio es crucial pero desafiante, especialmente para los ARN transcritos in vitro.
  • Los métodos existentes a menudo resultan en una modificación estocástica en grupos 2'-OH no emparejados con bases, lo que limita el control preciso.
  • La modificación de ARN localizada ofrece potencial para diversas aplicaciones, incluido el control de la función del ARN y la habilitación de etiquetas específicas.

Objetivo del estudio:

  • Desarrollar una nueva estrategia dirigida al ADN para la funcionalización selectiva del ARN en grupos 2'-OH.
  • Permitir un control preciso de la modificación del ARN para aplicaciones en biología molecular y biología química.
  • Para superar las limitaciones de los métodos estocásticos de modificación del ARN.

Principales métodos:

  • Desarrolló la acilación de ARN en bucles inducidos (RAIL), un método que utiliza oligonucleótidos de ADN complementarios.
  • Las sondas de ADN auxiliares crean huecos o bucles localizados en el ARN, exponiendo grupos 2'-OH específicos para la reacción.
  • Los reactivos de acilimidazol se utilizan para la conjugación de alto rendimiento en los sitios 2'-OH expuestos.

Principales resultados:

  • RAIL logra altos rendimientos de conjugación 2'-OH específica del sitio en moléculas de ARN.
  • Se identificaron diseños óptimos de oligodeoxinucleótidos auxiliares y condiciones de reacción.
  • El método controló con éxito la actividad localizada de la ribozima y permitió el etiquetado fluorescente de doble color del ARN.

Conclusiones:

  • El enfoque RAIL proporciona una estrategia simple y novedosa para el etiquetado y control de ARN selectivo del sitio.
  • Este método es aplicable a ARN de diversas longitudes y orígenes, ofreciendo una amplia utilidad.
  • RAIL avanza en el campo de la funcionalización del ARN, permitiendo la manipulación precisa de las moléculas de ARN.