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La arquitectura molecular para la escisión de ARN guiado por ARN por Cas13a

Liang Liu¹, Xueyan Li², Jun Ma³

¹Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

|August 1, 2017

Resumen

La enzima CRISPR-Cas13a, una potente ribonucleasa guiada por ARN, se activa mediante la unión al ARN objetivo. Este estudio estructural revela cómo Cas13a divide el ARN, allanando el camino para las tecnologías de manipulación de ARN.

Área de la Ciencia:

Biología molecular
La bioquímica
La genética

Sus antecedentes:

CRISPR-Cas13a es una ribonucleasa guiada por ARN con potencial en tecnología de ARN.
Comprender su mecanismo de activación es crucial para aprovechar sus capacidades.

Objetivo del estudio:

Para aclarar la base estructural de la activación de Cas13a y la escisión del ARN.
Para investigar los cambios conformacionales en Cas13a al unirse al ARN objetivo.

Principales métodos:

Determinación de la estructura cristalina de Leptotrichia buccalis (Lbu) Cas13a unido al crARN y al ARN objetivo.
Microscopía cryoelectrónica (cryo-EM) del complejo LbuCas13a-crRNA.

Principales resultados:

El dúplex crRNA-target se une dentro del canal central del lóbulo de la nucleasa.
La unión al ARN objetivo induce cambios conformacionales significativos en Cas13a.
La formación del dúplex de ARN guía-objetivo activa los dominios catalíticos HEPN para la escisión del ARN.

Conclusiones:

La activación de Cas13a implica un cambio conformacional desencadenado por la formación de duplex de ARN guía-objetivo.
La enzima activada divide tanto el ARN objetivo como el colateral.
Estos hallazgos iluminan el mecanismo de defensa de los sistemas CRISPR-Cas de tipo VI contra los fagos de ARN e informan el desarrollo de herramientas de manipulación de ARN.

Palabras clave:

C2c2 CRISPR-Cas también incluye:En el caso de los Escisión de ARN guiada por ARN La RNase

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