CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos

Hernandez-Huertas, L.; Diaz-Moscoso, A.; Brannan, E.O.; Bazzini, A.A.; Wang, W.; Monges, D.E.; Moreno-Sanchez, I.; Theune, W.C.; Kushawah, G.; Tomas-Gallardo, L.; Málaga Trillo, George Edward; Moran, A.M.; Sánchez Alvarado, A.; Abugattas-Nuñez del Prado, J.; Corbin, T.J.; Martinez-Morales, J.R.; Takacs, C.M.; Guelfo, J.R.; DeVore, M.L.; Hassan, H.; Moreno-Mateos, M.A.

dc.contributor.author	Kushawah, G.
dc.contributor.author	Hernandez-Huertas, L.
dc.contributor.author	Abugattas-Nuñez del Prado, J.
dc.contributor.author	Martinez-Morales, J.R.
dc.contributor.author	DeVore, M.L.
dc.contributor.author	Hassan, H.
dc.contributor.author	Moreno-Sanchez, I.
dc.contributor.author	Tomas-Gallardo, L.
dc.contributor.author	Diaz-Moscoso, A.
dc.contributor.author	Monges, D.E.
dc.contributor.author	Guelfo, J.R.
dc.contributor.author	Theune, W.C.
dc.contributor.author	Brannan, E.O.
dc.contributor.author	Wang, W.
dc.contributor.author	Corbin, T.J.
dc.contributor.author	Moran, A.M.
dc.contributor.author	Sánchez Alvarado, A.
dc.contributor.author	Málaga Trillo, George Edward
dc.contributor.author	Takacs, C.M.
dc.contributor.author	Bazzini, A.A.
dc.contributor.author	Moreno-Mateos, M.A.
dc.date.accessioned	2020-12-14T16:06:27Z
dc.date.available	2020-12-14T16:06:27Z
dc.date.issued	2020
dc.identifier.uri	https://hdl.handle.net/20.500.12866/8723
dc.description.abstract	The development of mRNA knockdown technologies for use in vertebrate organisms such as zebrafish has been limited. Kushawah et al. establish CRISPR-RfxCas13d as an efficient, specific, cost-effective, and straightforward method for the systematic and tractable study of gene function in vivo during embryogenesis across a range of animal species. © 2020 Elsevier Inc. 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.	en_US
dc.language.iso	eng
dc.publisher	Elsevier
dc.relation.ispartofseries	Developmental Cell
dc.rights	info:eu-repo/semantics/restrictedAccess
dc.rights.uri	https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject	CRISPR-Cas13	en_US
dc.subject	embryogenesis	en_US
dc.subject	zebrafish	en_US
dc.subject	RNA targeting	en_US
dc.subject	knockdown	en_US
dc.subject	early development	en_US
dc.subject	Cas13d	en_US
dc.subject	MZT	en_US
dc.subject	medaka	en_US
dc.subject	killifish	en_US
dc.title	CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos	en_US
dc.type	info:eu-repo/semantics/article
dc.identifier.doi	https://doi.org/10.1016/j.devcel.2020.07.013
dc.subject.ocde	https://purl.org/pe-repo/ocde/ford#1.06.01
dc.relation.issn	1878-1551