Engineering self-deliverable ribonucleoproteins for genome editing in the brain

The delivery of CRISPR ribonucleoproteins (RNPs) for genome editing in vitro and in vivo has important advantages over other delivery methods, including reduced off-target and immunogenic effects. However, effective delivery of RNPs remains challenging in certain cell types due to low efficiency and cell toxicity. To address these issues, we engineered self-deliverable RNPs that can promote efficient cellular uptake and carry out robust genome editing without the need for helper materials or biomolecules. Screening of cell-penetrating peptides (CPPs) fused to CRISPR-Cas9 protein identified potent constructs capable of efficient genome editing of neural progenitor cells. Further engineering of these fusion proteins identified a C-terminal Cas9 fusion with three copies of A22p, a peptide derived from human semaphorin-3a, that exhibited substantially improved editing efficacy compared to other constructs. We found that self-deliverable Cas9 RNPs generated robust genome edits in clinically ..., , , # Engineering self-deliverable ribonucleoproteins for genome editing in the brain

This dataset contains files of protein sequences, DNA sequences, RNA sequences, sequencing files, brain slice images, and flow analysis, used for the paper.

## Description of the data and file structure

The file entitled with “Protein_and_RNA_sequences” contains the RNA and protein sequences in the study.
Files entitled with “CL7_NLS_Cas9_A22p3-His”, “His_CL7_2NLS_iCas12a-2NLS”, “His_CL7_2NLS_iCas12a-4NLS”, “His_CL7_NLS_Cas9(dS)*A22p3”, “His_CL7_NLS_Cas9_A22p”, “His_CL7_NLS_Cas9_A22p3” are the snapgene files for the plasmid sequences in the study.
File entitled with “mGluR5-TH-NPC_and_mouse_brain_editing-NGS_data” contains the unprocessed next-generation sequencing data in the study.
File entitled with “Examples_of_flow_analysis” contains examples of flow analysis in the study.
Files entitled with “Brain_imaging*(tdTomato_Ai9)” and “Ai9_-_co-delivery_of_tdTom_and_TH_RNPs” contains the fluorescent imag...