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Edición continua asistida por helicasa para la mutagénesis programable de genomas endógenos

Xi Dawn Chen^1,2,3, Zeyu Chen^1,4,5, George Wythes¹

¹Gene Regulation Observatory, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.

Science (New York, N.Y.)

|October 10, 2024

Resumen

Una nueva herramienta llamada edición continua asistida por helicasa (HACE) permite mutaciones de ADN de largo alcance dirigidas. Esta plataforma ayuda a los investigadores a comprender la función de los genes y a descubrir nuevas capacidades biológicas.

Área de la Ciencia:

La genómica
Biología molecular
Ingeniería genética

Sus antecedentes:

Comprender las relaciones de secuencia-función en los genomas es crucial pero desafiante.
Los métodos actuales para la mutación y evolución genómica dirigida son limitados en alcance y aplicación.
El desarrollo de nuevas herramientas es esencial para diseccionar mecanismos genéticos complejos.

Objetivo del estudio:

Introducir una plataforma programable para la hipermutación genómica específica de largo alcance.
Para demostrar la utilidad de esta plataforma en los estudios de genómica funcional.
Permitir la investigación de los elementos genéticos codificantes y no codificantes.

Principales métodos:

Desarrollo de una plataforma de edición continua asistida por helicasa (HACE).
Utilizando CRISPR-Cas9 para guiar una enzima de fusión helicasa-deaminasa.
Aplicación de HACE para la mutagénesis dirigida a través de grandes intervalos genómicos (> 1000 bp).

Principales resultados:

HACE indujo con éxito la hipermutación en loci genómicos específicos.
Se han identificado mutaciones en MEK1 que confieren resistencia a los inhibidores de la quinasa.
Se diseccionó el impacto de las variantes en los errores dependientes de SF3B1.
Variantes no codificantes evaluadas dentro de un potenciador inmune CD69.

Conclusiones:

HACE es una herramienta poderosa para investigar las variantes codificadas y no codificadas.
La plataforma facilita el descubrimiento de las relaciones de secuencia-función combinatorias.
HACE permite la evolución dirigida de nuevas funciones biológicas.

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