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 vehicle..., , # Data from: Engineered nucleocytosolic vehicles for loading of programmable editors

\[10.1016/j.cell.2025.03.015]

Data collection for raw sequencing data, raw imaging files, and other data associated with the corresponding publication.

## Description of the data and file structure

* \"2Photon_in_vivo_imaging.zip\" contains the raw data of retinal images associated with the results presented in figure 7 and S14 in the manuscript. There are subfolders for the raw images generated for each eye explored in the manuscript, the folders are divided into ENVLPE and subsequent include every treated eye with extensive imaging from the respective area of the retina, both in black and white and additional the colored images.
\"ENVLP.zip\" file group contains the images of subject F (female), NP (no punch, mouse identification), injected with 1.5uL in the right eye (OD) with individual colored and black and white image reconstruction of the eye. Each of the individual images extracted from portio...,