PUBLICATION

Disease modeling by efficient genome editing using a near PAM-less base editor in vivo

Authors
Rosello, M., Serafini, M., Mignani, L., Finazzi, D., Giovannangeli, C., Mione, M.C., Concordet, J.P., Del Bene, F.
ID
ZDB-PUB-220616-3
Date
2022
Source
Nature communications   13: 3435 (Journal)
Registered Authors
Del Bene, Filippo, Mione, Marina, Rosello, Marion
Keywords
none
MeSH Terms
  • Animals
  • CRISPR-Associated Protein 9*/metabolism
  • CRISPR-Cas Systems/genetics
  • Gene Editing*/methods
  • Genome/genetics
  • Zebrafish/genetics
  • Zebrafish/metabolism
PubMed
35701478 Full text @ Nat. Commun.
Abstract
Base Editors are emerging as an innovative technology to introduce point mutations in complex genomes. So far, the requirement of an NGG Protospacer Adjacent Motif (PAM) at a suitable position often limits the base editing possibility to model human pathological mutations in animals. Here we show that, using the CBE4max-SpRY variant recognizing nearly all PAM sequences, we could introduce point mutations for the first time in an animal model with high efficiency, thus drastically increasing the base editing possibilities. With this near PAM-less base editor we could simultaneously mutate several genes and we developed a co-selection method to identify the most edited embryos based on a simple visual screening. Finally, we apply our method to create a zebrafish model for melanoma predisposition based on the simultaneous base editing of multiple genes. Altogether, our results considerably expand the Base Editor application to introduce human disease-causing mutations in zebrafish.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping