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Registro analógico reescribible de eventos múltiples en células bacterianas y de mamíferos
1Merkin Institute for Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA, and Department of Chemistry and Chemical Biology and Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02138, USA.
Science (New York, N.Y.)
|February 17, 2018
Resumen
Los científicos desarrollaron dos sistemas basados en CRISPR, CAMERA 1 y CAMERA 2, para registrar eventos celulares en bacterias y células de mamíferos. Estos "grabadores de datos celulares" almacenan permanentemente un historial de señales ambientales como modificaciones del ADN.
Área de la Ciencia:
- Biología sintética
- Biología molecular
- La genética
Sus antecedentes:
- Los procesos celulares son dinámicos e influenciados por numerosas señales endógenas y exógenas.
- Comprender las respuestas celulares requiere métodos para registrar con precisión los eventos pasados.
- Los métodos existentes para rastrear el historial celular son limitados en alcance y permanencia.
Objetivo del estudio:
- Desarrollar nuevos sistemas mediados por CRISPR para el registro de múltiples eventos celulares.
- Para crear sistemas de memoria basados en ADN para bacterias y células de mamíferos.
- Para permitir el registro de la amplitud, la duración y el orden de la señal.
Principales métodos:
- Utilizó editores de bases mediados por CRISPR y nucleasas Cas9.
- Desarrolló dos sistemas: CAMERA 1 (ratios de secuencia de ADN) y CAMERA 2 (modificaciones de una sola base).
- Sistemas diseñados para registrar diversos estímulos, incluidos antibióticos, nutrientes, virus, luz y señalización Wnt.
Principales resultados:
- Se registraron con éxito múltiples estímulos en células bacterianas y de mamíferos.
- Se ha demostrado una lectura confiable de tan solo 10-100 células cuando se utilizan plásmidos multicopia.
- Demostró la capacidad de registrar el orden de los estímulos y para borrar / volver a registrar recuerdos en múltiples ciclos.
Conclusiones:
- Los sistemas CAMERA funcionan con eficacia
- grabadoras de datos de las celdas
- escribiendo historias de eventos celulares en el ADN.
- Estos sistemas ofrecen un método permanente y de alta fidelidad para documentar la señalización celular.
- La tecnología tiene amplias aplicaciones en biología sintética, biología celular y comprensión de sistemas biológicos complejos.
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