期刊
DEVELOPMENTAL CELL
卷 54, 期 6, 页码 805-+出版社
CELL PRESS
DOI: 10.1016/j.devcel.2020.07.013
关键词
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资金
- Ramon y Cajal program from Spanish Ministerio de Ciencia, Innovacion y Universidades [RyC-2017-23041]
- Spanish Ministerio de Ciencia, Innovacion y Universidades [PGC2018-097260-B-I00, MDM-2016-0687, BFU2017-86339-P, MDM2016-0687]
- Springboard program from CABD
- Stowers Institute for Medical Research
- Genome Engineer Innovation 2019 Grant from Synthego
- Pew Innovation Fund
- INNOVATE PERU grant [168-PNICP-PIAP-2015]
- FONDECYT travel grant [043-2019]
- Pablo de Olavide University
- Consejo Superior de Investigaciones Cientificas (CSIC)
- Junta de Andalucia
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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