New prime editing method inserts DNA fragments up to 12.5 kilobases without double-strand breaks
Researchers at Westlake University developed donor-complementary prime editing (DoPE), a technique that inserts DNA sequences up to 12.5 kilobases into genomes without creating double-strand breaks. Published in Nature Biotechnology on 31 August 2026, the method combines a 3'-overhang double-stranded DNA donor with overhang-complementary prime editing guide RNAs and a PE2* prime editor. DoPE employing short (approximately 30-nucleotide) overhangs supports insertions ranging from small fragments to those exceeding 10 kilobases. The researchers replaced mutant exons of PRKCSH, either individually or simultaneously, to correct diverse mutations in vitro. The approach works in a single step, is compatible with pooled DNA libraries, and does not require recombinases or transposases.
The technique inserts large DNA fragments in a single step. It may advance genome editing in research and therapy.
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