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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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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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Interferencia de CRISPR en una cepa de tipo 17 de secuencia multilocal de Streptococcus agalactiae

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    Los investigadores desarrollaron un sistema de interferencia CRISPR para las cepas ST-17 del estreptococo del grupo B (GBS). Esta herramienta permite la eliminación de genes dirigidos para estudiar la patogénesis de la meningitis GBS en la barrera hematoencefálica.

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    Área de la Ciencia:

    • Microbiología
    • La genética
    • Enfermedades infecciosas

    Sus antecedentes:

    • El estreptococo del grupo B (GBS) es una de las principales causas de la meningitis bacteriana neonatal.
    • Las cepas hipervirulentas del serotipo III, tipo de secuencia 17 (ST-17), como la COH1, están fuertemente relacionadas con la enfermedad neonatal grave.
    • La manipulación genética de las cepas de ST-17 GBS es difícil, lo que dificulta la investigación de los factores de virulencia.

    Objetivo del estudio:

    • Desarrollar un sistema de interferencia CRISPR (CRISPRi) para la eliminación del gen objetivo en la cepa ST-17 GBS COH1.
    • Permitir la genómica funcional y el cribado de alto rendimiento de los factores de virulencia del SGB.
    • Para facilitar la investigación de la patogénesis del SGB en la barrera hematoencefálica.

    Principales métodos:

    • Desarrollo de un sistema de interferencia CRISPR (CRISPRi) utilizando Cas9 catalizador inactivado (dCas9) en la cepa COH1 GBS.
    • Confirmación de la eficacia del sistema mediante ensayos de hemólisis, qPCR y modelos de infección in vitro con células endoteliales del cerebro humano.
    • Eliminación dirigida de los genes de virulencia clave, incluidos pilA, srr2 y iagA.

    Principales resultados:

    • Implementación exitosa de un sistema CRISPRi sintonizable en el ST-17 GBS COH1.
    • Se ha demostrado la eliminación fenotípica de los genes esenciales de virulencia del SGB.
    • Se redujo la adhesión bacteriana, la invasión y las respuestas inflamatorias en la barrera hematoencefálica.

    Conclusiones:

    • El sistema CRISPRi desarrollado proporciona una plataforma versátil para la eliminación rápida de genes en ST-17 GBS.
    • Esta herramienta supera los desafíos anteriores de manipulación genética en COH1.
    • Permite la investigación avanzada sobre la patogénesis del SGB y las interacciones huésped-patógeno en la barrera hematoencefálica.