Safer and efficient base editing and prime editing via ribonucleoproteins delivered through optimized lipid-nanoparticle formulations

Delivering ribonucleoproteins (RNPs) for in vivo genome editing is safer than using viruses encoding for Cas9 and its respective guide RNA. However, transient RNP activity does not typically lead to optimal editing outcomes. Here, we show that the efficiency of delivering RNPs can be enhanced by cell-penetrating peptides (covalently fused to the protein or as excipients) and that lipid nanoparticles (LNPs) encapsulating RNPs can be optimized for enhanced RNP stability, delivery efficiency, and editing potency. Specifically, after screening for suitable ionizable cationic lipids and by optimizing the concentration of the synthetic lipid DMG-PEG 2000, we show that the encapsulation, via microfluidic mixing, of adenine base editor and prime editor RNPs within LNPs using the ionizable lipid SM102 can result in in vivo editing-efficiency enhancements larger than 300-fold (with respect to the delivery of the naked RNP) without detectable off-target edits. We believe that chemically defined LN..., , # Safer and efficient base editing and prime editing via ribonucleoproteins delivered through optimized lipid-nanoparticle formulations

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

## Description of the data and file structure

* The data are matched to figures and panels of the corresponding publication and sorted in the subdirectories. Below, X denotes a figure number.
* Fig X uncropped gels.pdf contain uncropped gels matched to Figure X, with appropriate panels assigned within the pdf file.
* FigX.csv contain numerical values used to prepare graphs, and the results of statistic tests as reported by GraphPad Prism 10 software.
* FigX_FC subdirectories contain flow cytometry data with events exported in .csv format and reports in .pdf format.
* FigX_FC_key.csv files contain a legend matching flow cytometry results to samples reported in the figures.
* Sequencing directory contains raw sequencing reads as dep...,

用于体内基因组编辑的核糖核蛋白（ribonucleoproteins, RNPs）递送策略，相较于编码Cas9及其对应向导RNA的病毒载体，安全性更优。然而，核糖核蛋白的瞬时活性通常无法实现最优的编辑效果。本研究证实，通过细胞穿透肽（cell-penetrating peptides，可与蛋白共价融合或作为辅料）可提升核糖核蛋白的递送效率；同时，可对包裹核糖核蛋白的脂质纳米颗粒（lipid nanoparticles, LNPs）进行优化，以增强核糖核蛋白的稳定性、递送效率与编辑效能。具体而言，在筛选合适的可电离阳离子脂质并优化合成脂质DMG-PEG 2000的浓度后，本研究表明，借助微流控混合技术，使用可电离脂质SM102将腺嘌呤碱基编辑器与先导编辑器核糖核蛋白包裹至脂质纳米颗粒中，可使体内编辑效率相较于裸核糖核蛋白递送提升300倍以上，且未检测到脱靶编辑。我们认为，化学定义的LN...

# 通过优化脂质纳米颗粒制剂递送核糖核蛋白实现安全高效的碱基编辑与先导编辑

本数据集包含与对应发表论文相关的原始测序文件、原始成像文件及其他各类数据。

## 数据与文件结构说明

* 本数据集与对应发表论文中的图表及子图一一匹配，并按子目录分类整理。下文以X指代图号。
* Fig X uncropped gels.pdf：包含与图X匹配的未裁切凝胶电泳图，PDF文件内已标注对应的子图分区。
* FigX.csv：包含用于绘制图表的数值数据，以及GraphPad Prism 10软件输出的统计检验结果。
* FigX_FC 子目录：包含流式细胞术数据，其中事件数据以.csv格式导出，分析报告以.pdf格式存储。
* FigX_FC_key.csv：包含流式细胞术结果与论文图表中样本的对应图例说明。
* Sequencing 目录：包含原始测序读段，具体信息详见原文完整内容。