GoKawiilNew 'Prime Assembly' method enables large-scale gene insertion using prime editing
Researchers used K562, Jurkat and HEK293T cell lines to test a genome-editing technique called prime assembly, which combines prime editing components (PE7 or PEmax) with pegRNAs and various DNA donor templates to insert genetic sequences at targeted genomic sites. The team tested donor formats ranging from short synthetic single-stranded DNA to large double-stranded DNA fragments up to 12.1 kilobases, optimizing nucleofection conditions for each donor type.
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This approach could expand prime editing beyond small sequence corrections toward inserting much larger genetic payloads directly into precise genomic locations, a capability with significant implications for gene therapy and cell engineering. The careful optimization of donor concentrations and delivery methods addresses a key practical barrier—cellular toxicity—that has limited the use of long DNA donors in precision genome editing.
- Prime assembly combines prime editing machinery with donor DNA templates to insert sequences at specific genomic locations.
- The method was tested across multiple donor DNA formats and sizes, from short ssDNA to 12.1-kb double-stranded DNA.
- Toxicity constraints required different concentration ranges depending on donor DNA length and structure.
Source: nature.com — Levesque, 2026-09-16
Published there as: “Targeted genomic integration and rearrangement using prime assembly”
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