CRISPR-Cas13d induces efficient mRNA knock-down in animal embryos

Kushawah, G.; Abugattas-Nunez del Prado, J.; Martinez-Morales, J. R.; DeVore, M.; Guelfo, J. R.; Brannan, E. O.; Wang, W.; Corbin, T. J.; Moran, A. M.; Sanchez Alvarado, A.; Malaga-Trillo, E.; Takacs, C. M.; Bazzini, A. A.; Moreno-Mateos, M. A.

doi:10.1101/2020.01.13.904763

bioRxiv · 10.1101/2020.01.13.904763

CRISPR-Cas13d induces efficient mRNA knock-down in animal embryos

Kushawah, G.·Abugattas-Nunez del Prado, J.·Martinez-Morales, J. R.·DeVore, M.·Guelfo, J. R.·Brannan, E. O.·Wang, W.·Corbin, T. J.·Moran, A. M.·Sanchez Alvarado, A.·Malaga-Trillo, E.·Takacs, C. M.·Bazzini, A. A.·Moreno-Mateos, M. A.

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Abstract

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 induce RNA degradation 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-Cas13d is an effective and precise system to deplete specific mRNA transcripts in zebrafish embryos. We demonstrate that both zygotically-expressed and maternally-provided transcripts are efficiently targeted, resulting in an 80% average decrease in transcript level and the 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-Cas13d is an efficient knock-down platform to interrogate gene function in animal embryos.

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Kushawah, G., Abugattas-Nunez del Prado, J., Martinez-Morales, J. R., DeVore, M., Guelfo, J. R., Brannan, E. O., Wang, W., Corbin, T. J., Moran, A. M., Sanchez Alvarado, A., Malaga-Trillo, E., Takacs, C. M., Bazzini, A. A., Moreno-Mateos, M. A.. 2020-01-14. CRISPR-Cas13d induces efficient mRNA knock-down in animal embryos. https://doi.org/10.1101/2020.01.13.904763

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