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Codificación de barras del desarrollo de ratones enteros a través de CRISPR

Reza Kalhor^1,2, Kian Kalhor³, Leo Mejia⁴

¹Department of Genetics, Harvard Medical School, Boston, MA, USA. gchurch@genetics.med.harvard.edu kalhor@genetics.med.harvard.edu.

Science (New York, N.Y.)

|August 11, 2018

Resumen

Desarrollamos un nuevo sistema de código de barras in vivo en ratones usando tecnología CRISPR para registrar la historia celular. Este método permite un seguimiento preciso del linaje y estudios de desarrollo en mamíferos.

Área de la Ciencia:

Biología del desarrollo
La genética
Biología molecular

Sus antecedentes:

El código de barras in vivo es crucial para comprender la información biológica, incluidos los linajes de desarrollo.
Las aplicaciones anteriores de codificación de barras de mutación inducida por nucleasa en sistemas de mamíferos han sido limitadas.
El rastreo de linaje basado en CRISPR ofrece potencial para el registro biológico de alta resolución.

Objetivo del estudio:

Establecer una plataforma versátil de código de barras in vivo en ratones.
Permitir el rastreo de linajes de alto rendimiento y estudios de desarrollo en mamíferos.
Para reconstruir los primeros linajes de desarrollo e investigar el desarrollo del eje cerebral.

Principales métodos:

Utilizó múltiples ARN guía de localización para la mutagénesis mediada por CRISPR in vivo.
Codificación de barras activada en el momento de la concepción y mutagénesis continua durante toda la gestación.
Generó códigos de barras exponencialmente diversos a través de mutaciones combinadas.
Código de barras aplicado para la reconstrucción post hoc de linajes tempranos y análisis del desarrollo cerebral.

Principales resultados:

Crearon con éxito ratones con código de barras con mutaciones específicas del linaje.
Logró una reconstrucción confiable de los primeros linajes de mamíferos.
Permitió la investigación de los procesos de desarrollo de la formación del eje cerebral.
Demostró la capacidad de la plataforma para el rastreo de linaje de alta resolución.

Conclusiones:

El sistema de código de barras desarrollado in vivo es una plataforma habilitadora y versátil para la investigación en mamíferos.
Esta tecnología avanza significativamente las capacidades de rastreo de linaje en modelos de ratón.
El sistema proporciona nuevas vías para estudiar la biología del desarrollo y los mecanismos de la enfermedad.

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