ZFIN ID: ZDB-PUB-200810-31
CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos
Kushawah, G., Hernandez-Huertas, L., Abugattas-Nuñez Del Prado, J., 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., Málaga-Trillo, E., Takacs, C.M., Bazzini, A.A., Moreno-Mateos, M.A.
Date: 2020
Source: Developmental Cell   54(6): 805-817.e7 (Journal)
Registered Authors: Málaga-Trillo, Edward, Martinez-Morales, Juan R., Takacs, Carter M.
Keywords: CRISPR-Cas13, Cas13d, MZT, RNA targeting, early development, embryogenesis, killifish, knockdown, medaka, zebrafish
MeSH Terms:
  • Animals
  • CRISPR-Cas Systems/genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics*
  • Gene Editing*/methods
  • Gene Expression Regulation, Developmental/genetics*
  • HEK293 Cells
  • Humans
  • RNA Interference/physiology
  • RNA, Messenger/genetics
PubMed: 32768421 Full text @ Dev. Cell
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.