Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain |
NC No Ligand Female |
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 |
15000001315 |
|
sNLS-SpCas9-sNLS |
RNP-NC-no ligand |
Nanoparticle |
Ai14 gRNA |
|
|
Mouse |
B6.Cg-Gt(ROSA)26Sor^tm14(CAG-tdTomato)Hze/J |
|
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 |
13-0300Â |
Thermo-Fisher, Rat anti-GFAP |
|
AB_10711040 |
ab104224Â |
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 |
28.87  |
1 |
25.0  |
2 |
27.6  |
3 |
34.0  |