PUBLICATION
Enhanced RNA-targeting CRISPR-Cas technology in zebrafish
- Authors
- Moreno-Sánchez, I., Hernández-Huertas, L., Nahón-Cano, D., Martínez-García, P.M., Treichel, A.J., Gómez-Marin, C., Tomás-Gallardo, L., da Silva Pescador, G., Kushawah, G., Egidy, R., Perera, A., Díaz-Moscoso, A., Cano-Ruiz, A., Walker, J.A., Muñoz, M.J., Holden, K., Galcerán, J., Nieto, M.Á., Bazzini, A.A., Moreno-Mateos, M.A.
- ID
- ZDB-PUB-250318-3
- Date
- 2025
- Source
- Nature communications 16: 25912591 (Journal)
- Registered Authors
- Keywords
- none
- Datasets
- GEO:GSE270724, GEO:GSE270530, GEO:GSE270527, GEO:GSE270528, GEO:GSE284120, GEO:GSE270529
- MeSH Terms
-
- RNA, Guide, CRISPR-Cas Systems*/genetics
- Zebrafish*/genetics
- Embryo, Nonmammalian/metabolism
- CRISPR-Cas Systems*
- Gene Editing/methods
- RNA/genetics
- RNA/metabolism
- Animals
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- PubMed
- 40091120 Full text @ Nat. Commun.
Citation
Moreno-Sánchez, I., Hernández-Huertas, L., Nahón-Cano, D., Martínez-García, P.M., Treichel, A.J., Gómez-Marin, C., Tomás-Gallardo, L., da Silva Pescador, G., Kushawah, G., Egidy, R., Perera, A., Díaz-Moscoso, A., Cano-Ruiz, A., Walker, J.A., Muñoz, M.J., Holden, K., Galcerán, J., Nieto, M.Á., Bazzini, A.A., Moreno-Mateos, M.A. (2025) Enhanced RNA-targeting CRISPR-Cas technology in zebrafish. Nature communications. 16:25912591.
Abstract
CRISPR-Cas13 RNA-targeting systems are widely used in basic and applied sciences. However, its application has recently generated controversy due to collateral activity in mammalian cells and mouse models. Moreover, its competence could be improved in vivo. Here, we optimized transient formulations as ribonucleoprotein complexes or mRNA-gRNA combinations to enhance the CRISPR-RfxCas13d system in zebrafish. We i) use chemically modified gRNAs to allow more penetrant loss-of-function phenotypes, ii) improve nuclear RNA targeting, and iii) compare different computational models and determine the most accurate to predict gRNA activity in vivo. Furthermore, we demonstrate that transient CRISPR-RfxCas13d can effectively deplete endogenous mRNAs in zebrafish embryos without inducing collateral effects, except when targeting extremely abundant and ectopic RNAs. Finally, we implement alternative RNA-targeting CRISPR-Cas systems such as CRISPR-Cas7-11 and CRISPR-DjCas13d. Altogether, these findings contribute to CRISPR-Cas technology optimization for RNA targeting in zebrafish through transient approaches and assist in the progression of in vivo applications.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping