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The Antiviral System of Bacteria and Archaea: CRISPR01:23

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Captura de ADN extraño durante la inmunidad adaptativa CRISPR-Cas

James K Nuñez1, Lucas B Harrington1, Philip J Kranzusch1,2

  • 1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California 94720, USA.

Nature
|October 28, 2015
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Resumen

Las bacterias y las arqueas utilizan la inmunidad CRISPR para defenderse de los virus mediante la integración de ADN extraño. Los investigadores revelaron cómo el complejo de enzimas Cas1-Cas2 captura el ADN, actuando como un regulador molecular para la construcción del locus CRISPR.

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Área de la Ciencia:

  • Biología molecular
  • Genética microbiana
  • Biología estructural

Sus antecedentes:

  • Las bacterias y las arqueas poseen sistemas inmunológicos adaptativos contra los fagos y los plásmidos.
  • Esta inmunidad implica la integración de fragmentos de ADN extraños (espaciadores) en los loci de CRISPR.
  • El complejo de integrasa Cas1-Cas2 es crucial para la adquisición de estos espaciadores.

Objetivo del estudio:

  • Elucidar el mecanismo por el cual el complejo Cas1-Cas2 selecciona sustratos de ADN extraños.
  • Comprender la base estructural de la adquisición de espaciadores en la inmunidad CRISPR.
  • Para revelar cómo Cas1-Cas2 funciona como un jefe molecular en la formación del locus CRISPR.

Principales métodos:

  • Se utilizó la cristalografía de rayos X para determinar la estructura del complejo Cas1-Cas2 de Escherichia coli.
  • El estudio incluyó el análisis del complejo ligado a sustratos de ADN de protospacer de 33 nucleótidos.
  • Los datos estructurales se interpretaron para comprender las interacciones proteína-ADN y la selección del sustrato.

Principales resultados:

  • El complejo Cas1-Cas2 forma una superficie de unión curva que abarca toda la longitud del ADN protospacer.
  • El complejo dispersa los extremos del ADN, posicionando los grupos 3'-OH terminales para el ataque nucleofílico dentro de los sitios activos de Cas1.
  • Las interacciones de la columna vertebral del fosfodiéster entre el ADN y el complejo proteico explican la selección de sustrato no específica de la secuencia observada.

Conclusiones:

  • El estudio revela la base estructural para la captura de ADN extraño por el complejo Cas1-Cas2.
  • Descubre el mecanismo por el cual Cas1-Cas2 actúa como una regla molecular, dictando el tamaño y la arquitectura de los loci de CRISPR.
  • Estos hallazgos proporcionan información crítica sobre el proceso fundamental de la inmunidad adaptativa en los procariotas.