Title
CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos
Date Issued
28 September 2020
Access level
open access
Resource Type
journal article
Author(s)
Kushawah G.
Hernandez-Huertas L.
Martinez-Morales J.R.
DeVore M.L.
Hassan H.
Moreno-Sanchez I.
Tomas-Gallardo L.
Diaz-Moscoso A.
Monges D.E.
Guelfo J.R.
Theune W.C.
Brannan E.O.
Wang W.
Corbin T.J.
Moran A.M.
Sánchez Alvarado A.
Takacs C.M.
Bazzini A.A.
Moreno-Mateos M.A.
Publisher(s)
Cell Press
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.
Start page
805
End page
817.e7
Volume
54
Issue
6
Language
English
OCDE Knowledge area
Zoología, Ornitología, Entomología, ciencias biológicas del comportamiento
Genética, Herencia
Subjects
Scopus EID
2-s2.0-85089988712
PubMed ID
Source
Developmental Cell
ISSN of the container
15345807
Sponsor(s)
We thank Raktim Roy, Kai Zhang, and Hui Qian for initial support; Fish facility, Molecular Biology facility, and Shiyuan Chen from the Computational core facilities from Stowers Institute; and Ana Fernández-Miñan from the CABD genomic and Genetics section within the Department of Molecular Biology and Biochemical Engineer at University Pablo de Olavide for reagents and experimental support. We also thank Paul Trainor and Michael Durnin for their help with mouse experiments, Mark Miller and Philippe Noguera for illustrations and animations, and Juan P. Fernandez for critical reading of the manuscript. We thank Ignacio Maeso (CABD), Alfonso Fernandez-Alvarez (CABD), and all members of the Moreno-Mateos and the Bazzini laboratories for intellectual and technical support. This work was supported by Ramon y Cajal program ( RyC-2017-23041 ) and grants PGC2018-097260-B-I00 and MDM-2016-0687 from Spanish Ministerio de Ciencia, Innovación y Universidades and the Springboard program from CABD (M.A.M.-M.) and the Stowers Institute for Medical Research (A.A.B.). M.A.M.-M. was the recipient of the Genome Engineer Innovation 2019 Grant from Synthego . A.A.B. was awarded with Pew Innovation Fund. J.R.M.-M. is supported by BFU2017-86339-P and MDM-2016-0687 grants (Spanish Ministerio de Ciencia, Innovación y Universidades). E.M.-T. and J.A.-N.d.P. are supported by INNOVATE PERÚ grant 168-PNICP-PIAP-2015 and FONDECYT travel grant 043-2019 . The CABD is an institution funded by Pablo de Olavide University , Consejo Superior de Investigaciones Científicas (CSIC), and Junta de Andalucía .
We thank Raktim Roy, Kai Zhang, and Hui Qian for initial support; Fish facility, Molecular Biology facility, and Shiyuan Chen from the Computational core facilities from Stowers Institute; and Ana Fern?ndez-Mi?an from the CABD genomic and Genetics section within the Department of Molecular Biology and Biochemical Engineer at University Pablo de Olavide for reagents and experimental support. We also thank Paul Trainor and Michael Durnin for their help with mouse experiments, Mark Miller and Philippe Noguera for illustrations and animations, and Juan P. Fernandez for critical reading of the manuscript. We thank Ignacio Maeso (CABD), Alfonso Fernandez-Alvarez (CABD), and all members of the Moreno-Mateos and the Bazzini laboratories for intellectual and technical support. This work was supported by Ramon y Cajal program (RyC-2017-23041) and grants PGC2018-097260-B-I00 and MDM-2016-0687 from Spanish Ministerio de Ciencia, Innovaci?n y Universidades and the Springboard program from CABD (M.A.M.-M.) and the Stowers Institute for Medical Research (A.A.B.). M.A.M.-M. was the recipient of the Genome Engineer Innovation 2019 Grant from Synthego. A.A.B. was awarded with Pew Innovation Fund. J.R.M.-M. is supported by BFU2017-86339-P and MDM-2016-0687 grants (Spanish Ministerio de Ciencia, Innovaci?n y Universidades). E.M.-T. and J.A.-N.d.P. are supported by INNOVATE PER? grant 168-PNICP-PIAP-2015 and FONDECYT travel grant 043-2019. The CABD is an institution funded by Pablo de Olavide University, Consejo Superior de Investigaciones Cient?ficas (CSIC), and Junta de Andaluc?a. M.A.M.-M. and A.A.B. conceived the project and designed the research. M.A.M.-M. G.K. A.A.B. L.H.-H. and J.A.-N.d.P. performed all zebrafish experiments with the contribution of M.L.D. I.M.-S. and J.R.G. W.C.T. and E.O.B. carried out CRISPR-LwaCas13a experiments and shh in situ experiments. L.T.-G. A.D.-M. and D.E.M. purified RfxCas13d protein. J.R.M.-M. M.A.M.-M. and J.A.-N.d.P. performed medaka experiments. W.W. T.J.C. A.M.M. and G.K. carried out killifish and mouse experiments. H.H. did primary RNA-seq analysis. E.M.-T. and A.S.A. provided reagents and materials. M.A.M.-M. A.A.B. and G.K. performed data analysis and M.A.M.-M. C.M.T. and A.A.B. wrote the manuscript with input from the other authors. All authors reviewed and approved the manuscript. The authors declare no competing interests.
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