Science Spotlight: New prime editing tools for large DNA insertions

Two groups showed that prime editing-derived platforms can integrate multi-kilobase cargos without double-strand breaks (DSBs), which avoids DSB-associated chromosomal rearrangements seen with nuclease-based knockin methods. The two techniques differ in how they generate and use donor DNA.

A team from the Chinese Academy of Sciences revealed its method, dubbed “PRIME‑In” in Nature Biomedical Engineering. The technique uses prime editing to create short microhomology between the genomic target and a plasmid donor, then uses paired genomic nicks to drive DSB-free knockin of 2-9.6 kb cargos. The approach achieved up to 85% efficiency in cell lines, including about 50% non-viral integration of 3 kb CAR constructs in primary human T cells...