While zebrafish is emerging as a new model system to study human diseases, an efficient methodology to generate precise point mutations at high efficiency is still lacking. Here we show that base editors can generate C-to-T point mutations with high efficiencies without other unwanted on-target mutations. In addition, we established a new editor variant recognizing an NAA PAM, expanding the base editing possibilities in zebrafish. Using these approaches, we first generated a base change in the ctnnb1 gene, mimicking oncogenic mutations of the human gene known to result in constitutive activation of endogenous Wnt signaling. Additionally, we precisely targeted several cancer-associated genes including cbl. With this last target we created a new zebrafish dwarfism model. Together our findings expand the potential of zebrafish as a model system allowing new approaches for the endogenous modulation of cell signaling pathways and the generation of precise models of human genetic disease associated-mutations.

Precise base editing for the in vivo study of developmental signaling and human pathologies in zebrafish / Rosello, M.; Vougny, J.; Czarny, F.; Mione, M. C.; Concordet, J. -P.; Albadri, S.; Del Bene, F.. - In: ELIFE. - ISSN 2050-084X. - 10:(2021), pp. e6555201-e6555215. [10.7554/eLife.65552]

Precise base editing for the in vivo study of developmental signaling and human pathologies in zebrafish

Mione M. C.;
2021-01-01

Abstract

While zebrafish is emerging as a new model system to study human diseases, an efficient methodology to generate precise point mutations at high efficiency is still lacking. Here we show that base editors can generate C-to-T point mutations with high efficiencies without other unwanted on-target mutations. In addition, we established a new editor variant recognizing an NAA PAM, expanding the base editing possibilities in zebrafish. Using these approaches, we first generated a base change in the ctnnb1 gene, mimicking oncogenic mutations of the human gene known to result in constitutive activation of endogenous Wnt signaling. Additionally, we precisely targeted several cancer-associated genes including cbl. With this last target we created a new zebrafish dwarfism model. Together our findings expand the potential of zebrafish as a model system allowing new approaches for the endogenous modulation of cell signaling pathways and the generation of precise models of human genetic disease associated-mutations.
2021
Rosello, M.; Vougny, J.; Czarny, F.; Mione, M. C.; Concordet, J. -P.; Albadri, S.; Del Bene, F.
Precise base editing for the in vivo study of developmental signaling and human pathologies in zebrafish / Rosello, M.; Vougny, J.; Czarny, F.; Mione, M. C.; Concordet, J. -P.; Albadri, S.; Del Bene, F.. - In: ELIFE. - ISSN 2050-084X. - 10:(2021), pp. e6555201-e6555215. [10.7554/eLife.65552]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11572/335159
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