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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
Published on: August 25, 2017
Control negativo de acción cis de la replicación del ADN en las células eucariotas
1Division of Basic Sciences, Fred Hutchinson Cancer Center, Seattle, Washington 98104.
Cell
|February 12, 1988
Resumen
La replicación de un episodio quimérico SV40-BPV ocurre una vez por ciclo celular, controlado por un mecanismo negativo de acción cis. Esto asegura el mantenimiento estable del episodio al prevenir la replicación dentro del mismo ciclo celular.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- La epigenética es la epigenética.
- Virología Virología.
Sus antecedentes:
- Las réplicas episómicas requieren un control preciso del ciclo celular para mantener un número de copias estable.
- Comprender los mecanismos que rigen la iniciación y terminación de la replicación del ADN es crucial para la estabilidad del genoma.
Objetivo del estudio:
- Para investigar el control de la replicación dependiente del ciclo celular de un virus simio quimérico, el virus del papiloma bovino 40 (SV40-BPV) fue replicado episómicamente.
- Identificar los mecanismos reguladores que aseguran la replicación una vez por ciclo celular del ADN episomal.
Principales métodos:
- Se utilizaron líneas celulares estables de mono que albergaban una réplica episomal quimérica SV40-BPV.
- Se analizó la frecuencia de replicación y el mantenimiento del número de copias del episodio a través de ciclos celulares.
- Investigó el papel del factor de iniciación del antígeno T en el control de la replicación.
Principales resultados:
- El replicón episomal SV40-BPV se replicó exactamente una vez por ciclo celular.
- El número de copias del episodio se mantuvo estable, incluso con un exceso de antígeno T.
- Se identificó un mecanismo de control negativo de acción cis, que previene la replicación de episodios duplicados.
Conclusiones:
- El control de la replicación de este episodio está mediado por un nuevo mecanismo negativo de acción cis, no por factores positivos limitantes.
- El antígeno SV40 T forma una asociación estable y potencialmente hereditaria con la plantilla episomal después de la replicación.
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