eLifeJul 7, 2026

Improved genome editing methods for precise DNA changes using prime editors in zebrafish

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗OA ↗

Updated Jul 8, 2026

Abstract

Up to 27.3% precise insertion was achieved using a nuclease-based Prime Editor in the zebrafish genome.

CRISPR/Cas9 technologies face challenges due to random insertions and deletions that limit precise genome manipulation.
Prime Editors offer a method for programmed integration of short DNA modifications into genomes.
Nickase-based Prime Editor (PE2) resulted in a higher ratio of precise edits compared to total edits.
Nuclease-based Prime Editor (PEn) demonstrated greater efficiency for short DNA modifications.
Gene modifications made using this approach were successfully transmitted to the next generation.
The method allows for genome editing without the need for additional donor DNA.

Simplified

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.

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Competing interests

YO, ML, AK, FB, AB, JB, CH, CT, SS No competing interests declared, MP, EG, SR, MB, MM employee of AstraZeneca

PubMed

Improved genome editing methods for precise DNA changes using prime editors in zebrafish

Abstract

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Funding

Competing interests

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Abstract

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Funding

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Related papers

You found one interesting study. We’ll send the next 7.

What Lands in Your Inbox Each Week:

Recent issues from the crispr gene editing brief