Precise gene editing in human cells using CRISPR-Cas12a and circular RNA

Editing frequencies of up to 40.75% were achieved using the split nickase-dependent circular RNA-mediated prime editor (sniCPE) in human cells.

• The smaller Cas12a protein enables the development of four new circular RNA-mediated prime editor systems.
• These systems can specifically target T-rich genomic regions, offering advantages over traditional Cas9-based methods.
• Nuclease-dependent systems achieved editing efficiencies of up to 10.42%, while niCPE and sniCPE reached up to 24.89% and 40.75% respectively without positive selection.
• One derivative system, called one-sniCPE, integrates all three RNA editing components under a single promoter.
• Editing of multiple genes simultaneously was demonstrated, albeit at low efficiency, using the nickase prime editors.

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