Data from: Engineered nucleocytosolic vehicles for loading of programmable editors

Advanced gene editing methods have accelerated biomedical discovery and
hold great therapeutic promise, but safe and efficient delivery of gene
editors remains challenging. In this study, we present a virus-like
particle (VLP) system featuring nucleocytosolic shuttling vehicles that
retrieve pre-assembled Cas-effectors via aptamer-tagged guide RNAs. This
approach ensures preferential loading of fully assembled editor
ribonucleoproteins (RNPs) and enhances the efficacy of prime editing, base
editing, trans-activators, and nuclease activity coupled to
homology-directed repair in multiple immortalized, primary, stem cell, and
stem-cell-derived cell types. We also achieve additional protection of
inherently unstable prime editing guide RNAs (pegRNAs) by shielding the
3'-exposed end with Csy4/Cas6f, further enhancing editing
performance. Furthermore, we identify a minimal set of packaging and
budding modules that can serve as a platform for bottom-up engineering of
enveloped delivery vehicles. Notably, our system demonstrates superior
per-VLP editing efficiency in primary T lymphocytes and two mouse models
of inherited retinal disease, highlighting its therapeutic potential.