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Iterative recombinase technologies for efficient and precise genome engineering across kilobase to megabase scales
Repeated DNA-cutting methods for accurate and efficient genome editing from small to large DNA regions
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Abstract
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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