| Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain |
| NC-CPP Males |
| 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. |
| striatum |
| neuron |
| Editing Efficiency |
| 15000001310 |
|
| sNLS-SpCas9-sNLS |
| RNP-NC-CPP |
| Nanoparticle |
| Ai14 gRNA |
|
|
| Mouse |
| B6.Cg-Gt(ROSA)26Sor^tm14(CAG-tdTomato)Hze/J |
|
| 8 weeks |
| Male |
|
| 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 |
| brain (striatum) |
|
| once |
| 0.2 ul per minute |
| 1 microliter per hemisphere |
| 14 days |
| Nanocage with CRISPR/Cas9-RNP |
|
|
|
| AB_2209751 |
| Rockland Cat# 600-401-379 |
| Anti-RFP (RABBIT) Antibody |
|
| AB_2532994 |
| 13-0300 |
| Thermo-Fisher, Rat anti-GFAP |
|
| AB_10711040 |
| ab104224 |
| Abcam, mouse anti-NeuN |
|
| AB_141637 |
| A21207 |
| Invitrogen, Donkey anti-rabbit alexa fluor 594 |
|
| AB_2813835 |
| ab150155 |
| Abcam, Donkey anti-rat alexa fluour 647 |
|
| AB_141607 |
| A21202 |
| Invitrogen, Donkey anti-mouse alexafluor 488 |
|
| Measured Values |
| Samples Size: | 3 |
| |
| Mean |
26.3  |
| 1 |
25.7  |
| 2 |
28.5  |
| 3 |
24.7  |