Targeted genomic integration and rearrangement using prime assembly.

Prime assembly achieves targeted integration and large-scale genomic rearrangements in human cells without relying on double-stranded DNA donors or cell cycle cues.

• Why it matters: Effective therapeutic genome editing requires precise insertion of medium-to-large DNA sequences, a challenge that limits treatment of many genetic disorders. Overcoming these barriers could significantly advance gene therapies.
• What they did: The researchers developed prime assembly, a method using CRISPR-targeted dual flap synthesis to assemble and insert DNA sequences, performing targeted exon recoding, transgene integration, and megabase-scale rearrangements in primary human cells.
• The result: This approach broadens genome engineering capabilities by enabling site-specific integration of large DNA fragments in both dividing and non-dividing cells, paving the way for improved therapeutic interventions.