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Fanzor es una endonucleasa eucariota programable guiada por ARN

Makoto Saito^1,2,3,4,5, Peiyu Xu^1,2,3,4,5, Guilhem Faure^1,2,3,4,5

¹Broad Institute of MIT and Harvard, Cambridge, MA, USA.

|June 28, 2023

Resumen

Los eucariotas poseen endonucleasas guiadas por ARN, ejemplificadas por las proteínas Fanzor (Fz), que son cruciales para la ingeniería del genoma. Este descubrimiento confirma los sistemas guiados por ARN en los tres dominios de la vida.

Área de la Ciencia:

Biología molecular
La genética
La bioquímica

Sus antecedentes:

Los sistemas guiados por ARN, como CRISPR-Cas, son vitales en las procariotas para la inmunidad adaptativa y la escisión del ADN.
Los sistemas guiados por ARN eucariotas son menos conocidos, sin endonucleasas guiadas por ARN confirmadas.
Los sistemas OMEGA procariotas, incluido TnpB, comparten ascendencia con las proteínas Fanzor (Fz) eucariotas.

Objetivo del estudio:

Para caracterizar bioquímicamente las proteínas Fanzor (Fz).
Para investigar el potencial de Fz para la reprogramación en la ingeniería del genoma humano.
Para aclarar la base estructural de la actividad de la endonucleasa de ADN guiada por ARN de Fz.

Principales métodos:

Ensayos bioquímicos para determinar la actividad de la nucleasa de Fz.
Reprogramando Fz para la división del ADN en las células humanas.
Microscopía electrónica criogénica (cryo-EM) para el análisis estructural de Fz.

Principales resultados:

Las proteínas Fanzor (Fz) funcionan como endonucleasas de ADN guiadas por ARN.
Fz puede ser diseñado con éxito para aplicaciones de edición de genoma.
El análisis estructural revela regiones conservadas entre Fz, TnpB y Cas12.

Conclusiones:

Fanzor (Fz) representa un sistema OMEGA eucariótico.
Las endonucleasas guiadas por ARN están presentes en los tres dominios de la vida (bacterias, arqueas y eucariotas).
Fz amplía el conjunto de herramientas para la ingeniería del genoma eucariota.

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