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Ribozymes02:47

Ribozymes

12.3K
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can...
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Restriction Enzymes01:11

Restriction Enzymes

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Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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.
The recognition sites for Cre recombinase called LoxP...
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Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
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RNA Interference01:23

RNA Interference

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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...
26.0K
Homologous Recombination02:31

Homologous Recombination

50.5K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Updated: Jul 4, 2025

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

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La ribozima endonucleolítica hidrolítica (HYER) es programable para la escisión de ADN específica de la secuencia

Zi-Xian Liu1, Shouyue Zhang1, Han-Zhou Zhu1

  • 1Beijing Advanced Innovation Center for Structural Biology, State Key Laboratory of Membrane Biology, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.

Science (New York, N.Y.)
|February 1, 2024
PubMed
Resumen

Los investigadores descubrieron ribosimas endonucleolíticas hidrolíticas (HYER), un tipo de ARN catalítico, que puede cortar el ADN. Estas ribozimas naturales muestran potencial para aplicaciones precisas de manipulación de ADN y edición de genoma.

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Área de la Ciencia:

  • Biología molecular
  • La bioquímica
  • La genética

Sus antecedentes:

  • Las ribozimas son moléculas catalíticas de ARN con funciones esenciales en los procesos celulares.
  • Las ribozimas naturales son cada vez más reconocidas por su potencial más allá de la catálisis del ARN, incluida la manipulación del ADN.

Objetivo del estudio:

  • Identificar y caracterizar las ribozimas naturales que funcionan como endonucleasas de ADN específicas de la secuencia.
  • Para explorar la reutilización de estas ribozimas como nuevas herramientas de manipulación de ADN.

Principales métodos:

  • Centrándose en los intrones bacterianos del grupo II-C, los investigadores examinaron los sistemas que carecen de proteínas codificadas por intrones.
  • Se utilizaron ensayos in vitro para comprobar la actividad de escisión de las ribozimas identificadas (HYER) frente a varios sustratos de ácido nucleico.
  • Se empleó microscopía criolectrónica para determinar la base estructural del mecanismo catalítico y de unión al ADN de HYER1.
  • Se aplicaron estrategias de diseño racional para diseñar variantes HYER con mayor especificidad y funcionalidad.

Principales resultados:

  • Se identificaron varios sistemas de intrones, denominados Ribozimas Endonucleolíticos Hidrolíticos (HYER), que dividen el ARN, el ADN de una sola cadena y el ADN de doble cadena (dsDNA).
  • HYER1 demostró la capacidad de inducir rupturas del ADN en genomas de mamíferos in vitro.
  • La microscopía criolectrónica reveló una estructura homodimérica de HYER1 con una bolsa de hidrólisis dependiente de Mg2+ y capacidad de unión al ADN.
  • Las variantes HYER diseñadas exhibieron una mejor especificidad y flexibilidad de manipulación de ADN.

Conclusiones:

  • Las ribozimas naturales, específicamente las HYER del grupo II-C, poseen actividad de la endonucleasa del ADN.
  • Estos HYER pueden ser diseñados para una escisión precisa del ADN, ofreciendo una nueva plataforma prometedora para la edición del genoma y la biotecnología.