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ExperimentIndependent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Experiment ConditionD10, RNP, Female
Assay Description: Editing efficiency in small (<50000 um2) airways such as bronchioles and large (>50000 um2) airways such as the main bronchus, calculated by counting fluorescent cells (GFP) compared to DAPI stained cells.
Tissue Measured:uterus
Measurement Type:Editing Efficiency
Record ID15000001187

EditorSpCas9
Delivery SystemD10
Delivery System SubtypeAmphiphilic peptide
GuideSpyCas9 g-loxP2_C9
Vector

Model SpeciesMouse
Model NameB6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J

Application Methodintranasal
Application Siteairway epithelium
Dosage40 micromolar peptide, 2.5 micromolar SpCas9, 2 micromolar RNA
Injection Frequencytwice, one day apart
Injection Rate
Injection Volume50 microliters
Days post injection1 week
Editor Formatribonucleoprotein (RNP)
Antidote Id
Antidote Description

Measured Values
Samples Size: 6
 
Replicate Result (Editing Efficiency)
Measurement Units: Signal
Mean No editing detected 
1 No editing detected 
  

Title Description File Download SCGE ID
Heaney-SATC_McCray-Validation_Lung Inflation and Fixation Protocol Procedure for mouse lung inflation and fixation. Heaney-SATC_McCray-Validation_Lung_Inflation_and_Fixation.pdf 21000000022
Heaney-SATC_McCray-Validation_Intranasal Instillation in Mice Protocol Procedure for intranasal instillation of ribonucleoprotein/peptide complex in mice for delivery to the lung. Heaney-SATC_McCray-Validation_Intranasal_Instillation_in_Mice.pdf 21000000021
Heaney_SATC Tissue Processing, Imaging and Analysis Procedure for tissue preparation, imaging and analysis. Heaney_SATC_Tissue_Processing_Imaging_and_Analysis.pdf 21000000025