Modular lipid nanoparticle delivery of base editor components enables versatile platform strategies for therapeutic genome editing

Engineering next-generation cell therapies, from hypoimmunogenic iPSCs to allogeneic CAR-T cells, requires simultaneous execution of multiple complex edits: targeted transgene integration, multi-gene knockout, and preservation of genomic integrity. Conventional nuclease-based approaches can achieve this, but at a cost: double-strand breaks, chromosomal translocations, and imprecise off-target donor template integration.

In this ASGCT 2026 poster, Revvity presents a breakthrough application of the Pin-point™ modular base editing platform - demonstrating how a single design insight enables simultaneous, precise transgene knock-in and multiplexed base editing without the risks associated with nuclease editing.

The key innovation lies in decoupling deaminase recruitment from transgene targeting, allowing the same editing reaction to independently perform precise transgene integration and multiplexed base editing.

By designing sgRNA components with and without aptamers within the same editing reaction, the nCas nickase component can be independently directed to:

Transgene integration sites, using aptamer-less sgRNAs that generate paired nicks for HDR without recruiting the deaminase Base editing targets, using aptamer-encoding sgRNAs that recruit the deaminase for precise C>T or A>G conversion

Key findings:

• Hypoimmunogenic iPSCs engineered with simultaneous HLA-E knock-in and B2M/CIITA knockout maintain genomic integrity, confirmed by Optical Genome Mapping showing near-zero structural variants in edited clones vs. 86 in the parental line
• Single-copy, precise transgene integration confirmed by ddPCR and site-specific knock-in at the B2M locus (Chr15)
• Allogeneic CAR-T cells manufactured in a single intervention, simultaneous CD19 CAR integration via AAV donor + knockout of HLA-1, CD52, TCRα/β, and PD1
• Near-zero chromosomal translocations with the Pin-point platform vs. SpCas9 nuclease (targeted hybridisation-capture sequencing)
• Undetectable off-target AAV donor integration at sgRNA-targeted sites, a critical safety advantage over nuclease editing
• Cell viability and pluripotency preserved in iPSCs across multiplexed editing conditions, in stark contrast to SpCas9, which markedly reduced live cell counts and Nanog expression

The Pin-point™ base editing platform technology is available for clinical or diagnostic study and commercialization under a commercial license from Revvity.​