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ExperimentEnabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
Experiment ConditionRNP-NC-CPP
Assay Description: Editing efficiency calculated by counting the percent of NeuN+ neurons expressing tdTomato surrounding the injected area of the striatum
Tissue Measured:cerebral cortex
Cell Type :neuron
Measurement Type:Editing Efficiency
Record ID15000000946

EditorsNLS-SpCas9-sNLS
Delivery SystemRNP-NC-CPP
Delivery System SubtypeNanoparticle
GuideAi14 gRNA
Vector

Model SpeciesMouse
Model NameB6;129S6-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J

Application Methodstereotaxic injection
Application Siteintracerebral/intrastriatal
Dosage30 pmol
Injection Frequencyonce bilaterally
Injection Rate0.2 μl/min
Injection Volume1.5μl
Days post injection14 days
Editor FormatRNP
Antidote Id
Antidote Description

Measured Values
Samples Size: 3
 
Replicate Result (Editing Efficiency)
Measurement Units: % of cells
Mean 17.4 
1 24.6 
2 12.4 
3 15.2 
  
NC-CPP delivery of CRISPR RNP produces neuronal genome editing in animal UW1

Representative coronal brain images from an Ai14 reporter mouse intrastriatally injected with cell penetrating peptide (CPP) decorated nanocages (NC-CPP) carrying CRISPR Cas9/sgRNA ribonucleoprotein. The brain sections are shown in rostrocaudal (top to bottom) anatomical order. The colors correspond to tdTomato-expressing, genome-edited cells (red; white arrows pointing to edited area) with NeuN+ neurons (green), GFAP+ astrocytes (pink), and DAPI labelled nuclei (blue).


Title Description File Download SCGE ID
Gong_Intracranial Injection Procedure for Mice Procedure for intracranial delivery to mouse brain. Gong_Intracerebral_Injection_Protocol.pdf 21000000020