Experiment Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain Experiment Condition NC No Ligand Male Assay Description: Editing efficiency calculated by counting the number of Edited Neurons (NeuN+/GFP+ and TdTomato+/RFP+) out of the total number of neurons (NeuN+/GFP+) in a defined region of interest in the brain, expressed as a percentage of neurons in the ROI. Tissue Measured: striatum Cell Type : neuron Measurement Type: Editing Efficiency Record ID 15000001314 Editor sNLS-SpCas9-sNLS Delivery System RNP-NC-no ligand Delivery System Subtype Nanoparticle Guide Ai14 gRNA Vector Model Species Mouse Model Name B6.Cg-Gt(ROSA)26Sor^tm14(CAG-tdTomato)Hze/J Application Method Stereotactic bilateral intra-striatal injection, coordinates: Ant: + 1.2 mm from bregma and M/L +/-1.6 mm from midline; Depth = 3.25/3.75 mm Application Site brain (striatum) Dosage Injection Frequency once Injection Rate 0.2 ul per minute Injection Volume 1 ul per hemisphere Days post injection 14 days Editor Format Nanocage with CRISPR/Cas9-RNP Antidote Id Antidote Description Antibody RRID AB_2209751 Antibody Other ID Rockland Cat# 600-401-379 Antibody Description Anti-RFP (RABBIT) Antibody Antibody RRID AB_2532994 Antibody Other ID 13-0300  Antibody Description Thermo-Fisher, Rat anti-GFAP Antibody RRID AB_10711040 Antibody Other ID ab104224  Antibody Description Abcam, mouse anti-NeuN Antibody RRID AB_141637 Antibody Other ID A21207 Antibody Description Invitrogen, Donkey anti-rabbit alexa fluor 594 Antibody RRID AB_2813835 Antibody Other ID ab150155 Antibody Description Abcam, Donkey anti-rat alexa fluour 647 Antibody RRID AB_141607 Antibody Other ID A21202 Antibody Description Invitrogen, Donkey anti-mouse alexafluor 488 Measured Values Samples Size: 3   Replicate Result (Editing Efficiency) Measurement Units: % cells Mean 22.0  1 16.7  2 30.4  3 18.9