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ExperimentTesting new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in adult mouse cochlea
Experiment Condition306-S10
Assay Description: Editing efficiency is calculated by percentage of tdTomato positive cells
Tissue Measured:base of cochlear canal
Cell Type :Deiter's cell
Measurement Type:Editing Efficiency
Record ID15000000598

EditorCleanCap® Cas9
Delivery System306-S10
Delivery System SubtypeNanoparticle
Guidesg298
Vector

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

Application Methodcochleostomy
Application Sitecochlea
DosageLNP/Cas9/sgRNA=0.15/0.01/0.01ug
Injection Frequencyonce
Injection Rate3 nl/sec
Injection Volume300 nanoliters
Days post injection5 days
Editor FormatmRNA
Antidote Id
Antidote Description

Antibody RRIDAB_10015251
Antibody Other ID25-6790
Antibody DescriptionAnti-Myosin VIIa antibody, Proteus Biosciences

Antibody RRIDAB_2286684
Antibody Other IDsc-17320
Antibody DescriptionSox-2 (Y-17) Antibody, Santa Cruz Biotechnology

Measured Values
Samples Size: 4
 
Replicate Result (Editing Efficiency)
Measurement Units: % of cells
Mean 17.36 
1 10.53 
2 25 
3 19.05 
4 14.85 
  
Delivery of Cas9/sgRNA mixed with 306-S10 into adult mouse cochlea via cochleostomy

Delivery of Cas9/sgRNA mixed with 306-S10 into adult mouse cochlea via cochleostomy. a-f. Whole mount immunofluorescence staining of the base region of the cochlea using MYO7A (green) and SOX2 (cyan). Cochlea tissues were collected 3-6 days post injection. tdT signals were observed mainly in Deiters cells. (a-b), low magnification images; (c-d), high magnification images. (e-f) cross section view of (c-d). G, illustration of cells types with tdT signal in cochlea. H, quantification of tdT+ cells in cochlea. N=2, mean +/- SEM. DC, Deiters cells; OPC, outer pillar cells; IHC, inner hair cells.