Nature communicationsDec 10, 2025

Programmable DNA integration using ligase enzymes

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

Updated Jun 27, 2026

Abstract

Essence

L-PGI is a ligase-based genome-editing method designed to insert synthetic DNA donors at Cas9 nickase sites without double-strand breaks.

Evidence

This preclinical platform study reports editing activity in cell lines, primary cell cultures, and adult mice after nonviral delivery.

Caveat

The evidence is limited to experimental models, and the abstract does not report durable disease correction or human therapeutic outcomes.

Simplified

Since their discovery, CRISPR systems have been repurposed for programmable targeted genomic editing, leading to applications for gene disruption, single base editing, insertion, deletion, and manipulation of short genomic sequences. Pairing Cas9 nickase with reverse transcriptase allows applications for insertion, substitution, and deletion of short genomic sequences from an RNA template without generating double stranded breaks however this technology typically shows reduced efficacy in post mitotic cells, limiting its translatability in vivo. Here we present a novel, ligase-based method that addresses these limiations. We introduce edits through delivery and ligation of a synthetic DNA donor to genomic nicks created with Cas9 nickase and report editing activity in cell lines, primary cell cultures, and adult mice via nonviral delivery. With favorable on target outcomes compared to transcription-based editing in key cell types, good tolerability, and deliverability, ligation-mediated gene editing has the potential to further advance genomic medicine.