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ExperimentIndependent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain
Experiment ConditionNC No Ligand Female
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 ID15000001315

EditorsNLS-SpCas9-sNLS
Delivery SystemRNP-NC-no ligand
Delivery System SubtypeNanoparticle
GuideAi14 gRNA
Vector

Model SpeciesMouse
Model NameB6.Cg-Gt(ROSA)26Sor^tm14(CAG-tdTomato)Hze/J

Application Methodstereotactic 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 Sitebrain (striatum)
Dosage
Injection Frequencyonce
Injection Rate0.2 ul per minute
Injection Volume1 microliter per hemisphere
Days post injection14 days
Editor FormatNanocage with CRISPR/Cas9-RNP
Antidote Id
Antidote Description

Antibody RRIDAB_2209751
Antibody Other IDRockland Cat# 600-401-379
Antibody DescriptionAnti-RFP (RABBIT) Antibody

Antibody RRIDAB_2532994
Antibody Other ID13-0300 
Antibody DescriptionThermo-Fisher, Rat anti-GFAP

Antibody RRIDAB_10711040
Antibody Other IDab104224 
Antibody DescriptionAbcam, mouse anti-NeuN

Antibody RRIDAB_141637
Antibody Other IDA21207
Antibody DescriptionInvitrogen, Donkey anti-rabbit alexa fluor 594

Antibody RRIDAB_2813835
Antibody Other IDab150155
Antibody DescriptionAbcam, Donkey anti-rat alexa fluour 647

Antibody RRIDAB_141607
Antibody Other IDA21202
Antibody DescriptionInvitrogen, Donkey anti-mouse alexafluor 488

Measured Values
Samples Size: 3
 
Replicate Result (Editing Efficiency)
Measurement Units: % cells
Mean 28.87 
1 25.0 
2 27.6 
3 34.0 
  

Title Description File Download SCGE ID
Murray-SATC_Gong-Validation Brain Injection in Mice Protocol Procedure for brain injection surgical procedure, pre- and post-operative care for mice. Murray-SATC_Gong-Validation_Brain_Injection_in_Mice.pdf 21000000026
Murray-SATC_Gong-Validaiton_Gong Study Protocol Procedure for intracranial injection, immunofluorescence and imaging. Murray-SATC_Gong-Validation_Gong_ Study_Protocol.pdf 21000000027