CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos

Early embryonic development is driven exclusively by maternal gene products deposited into the oocyte. Although critical in establishing early developmental programs, maternal gene functions have remained elusive due to a paucity of techniques for their systematic disruption and assessment. CRISPR-C...

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Detalles Bibliográficos
Autores: Kushawah, Gopal, Abugattas-Nuñez del Prado, Joaquin A., Martínez-Morales, Juan Ramón, DeVore, Michelle L., Hassan, Huzaifa, Moreno-Sánchez, Ismael, Tomás-Gallardo, Laura, Díaz Moscoso, Alejandro, Monges, Dahiana E., Guelfo, Javier R., Theune, William C., Brannan, Emry O., Wang, Wei, Corbin, Timothy J., Moran, Andrea M., Sánchez Alvarado, Alejandro, Málaga-Trillo, Edward, Takacs, Carter M., Bazzini, Ariel A., Moreno-Mateos, Miguel A.
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2020
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/226267
Acceso en línea:http://hdl.handle.net/10261/226267
Access Level:acceso abierto
Palabra clave:CRISPR-Cas13
Embryogenesis
Zebrafish
RNA targeting
Knockdown
Early development
Cas13d
MZT
Medaka
Killifish
Descripción
Sumario:Early embryonic development is driven exclusively by maternal gene products deposited into the oocyte. Although critical in establishing early developmental programs, maternal gene functions have remained elusive due to a paucity of techniques for their systematic disruption and assessment. CRISPR-Cas13 systems have recently been employed to degrade RNA in yeast, plants, and mammalian cell lines. However, no systematic study of the potential of Cas13 has been carried out in an animal system. Here, we show that CRISPR-RfxCas13d (CasRx) is an effective and precise system to deplete specific mRNA transcripts in zebrafish embryos. We demonstrate that zygotically expressed and maternally provided transcripts are efficiently targeted, resulting in a 76% average decrease in transcript levels and recapitulation of well-known embryonic phenotypes. Moreover, we show that this system can be used in medaka, killifish, and mouse embryos. Altogether, our results demonstrate that CRISPR-RfxCas13d is an efficient knockdown platform to interrogate gene function in animal embryos.

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