PUBLICATION
Optimised genome editing for precise DNA insertion and substitution using prime editors in zebrafish
- Authors
- Ono, Y., Peterka, M., Love, M., Khan, A., Bowers, F., Bhandari, A., Gordon, E., Ball, J.S., Hammond, C., Tyler, C.R., Rees, S., Bohlooly-Y, M., Maresca, M., Scholpp, S.
- ID
- ZDB-PUB-260708-1
- Date
- 2026
- Source
- eLIFE 14: RP107475 (Journal)
- Registered Authors
- Bhandari, Ashish, Hammond, Chrissy, Ono, Yosuke, Scholpp, Steffen, Tyler, Charles R.
- Keywords
- CRISPR/Cas9, PE2, PEn, genetics, genomics, precise genome editing, prime editing, zebrafish
- MeSH Terms
- none
- PubMed
- 42411453 Full text @ Elife
Citation
Ono, Y., Peterka, M., Love, M., Khan, A., Bowers, F., Bhandari, A., Gordon, E., Ball, J.S., Hammond, C., Tyler, C.R., Rees, S., Bohlooly-Y, M., Maresca, M., Scholpp, S. (2026) Optimised genome editing for precise DNA insertion and substitution using prime editors in zebrafish. eLIFE. 14:RP107475.
Abstract
CRISPR/Cas9-mediated genome editing has rapidly become a popular tool for studying gene functions and generating genetically modified organisms. However, using this system, stochastic integration of random insertions and deletions restricts precise genome manipulation. Advanced CRISPR/Cas9 technologies using Prime Editors (PEs), Cas9 proteins fused with reverse transcriptase, enable programmed integration of short DNA modifications into the genome. However, its application in precise genome editing in animal models is challenging. Here, we utilise a nickase- and a nuclease-based PE to perform programmed short DNA substitutions and insertions at various loci in the zebrafish genome. Whereas nickase-based PE2 mediated a higher ratio of precise prime edits to the total edits, nuclease-based PEn was more efficient for short DNA modifications, achieving up to 27.3% precise insertion. To further evaluate our approach, we inserted a nuclear localisation signal into a reporter transgene to incorporate longer fragments by prime editing. These gene modifications were transmitted to the next generation. We show that PE-mediated prime editing can efficiently manipulate genome information in zebrafish without using exogenous donor DNA.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping