Repeated DNA-cutting methods for accurate and efficient genome editing from small to large DNA regions

Programmable chromosome engineering (PCE) enables scarless kilobase-to-megabase DNA manipulations in plants and human cells.

• PCE and RePCE are new systems designed for precise chromosome editing.
• A 10-fold reduction in the reversibility of Lox sites was achieved through high-throughput engineering.
• AI-assisted recombinase engineering resulted in Cre variants with 3.5 times the recombination efficiency compared to the wild type.
• The scar-free strategy allows for insertions, deletions, replacements, inversions, and translocations at the chromosomal level.
• Applications include a 315-kb inversion in rice that provides herbicide resistance and a 12-Mb inversion at sites related to human diseases.

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