L1+R1, unmodified scaffold

Experiment	Selection of gRNA sequences and gRNA scaffold modification lead to improved editing of the Ai9 locus in vitro
Experiment Condition	L1+R1, unmodified scaffold
Assay Description:	Editing efficiency was assessed by flow cytometry for tdTomato expression in Ai9 mouse embryonic fibroblasts after transfection with a mixture of plasmids expressing SaCas9 and the indicated U6-gRNAs
Tissue Measured:
Measurement Type:	Editing Efficiency
Record ID	15000000567

Editor	SaCas9
Delivery System
Delivery System Subtype
Guide	L1-unmodified; R1-unmodified
Vector	AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1; AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1;

Model Species	Mouse
Model Name	Ai9 Immortalized Fibroblasts

Application Method	Transient transfection
Application Site
Dosage
Injection Frequency
Injection Rate
Injection Volume
Days post injection
Editor Format	plasmid
Antidote Id
Antidote Description

Measured Values
Samples Size:	3

Replicate	Result (Editing Efficiency)
Measurement Units:	% tdTomato+ cells
Mean	2.78
1	4.22
2	3.58
3	0.55

Title	Description	File Download	SCGE ID
Deverman_AAV Production and Administration Protocol	Published procedures for AAV production, delivery, and tissue imaging. Challis et al. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.	Deverman_AAV_production_and_administration_Nat Protocols-Challis_et_al_2019.pdf	21000000010
Deverman_Comprehensive Methods	Procedure for plasmid cloning, editing evaluation in fibroblast, AAV production and administration, tissue processing and IHC.	Deverman_Comprehensive_methods.pdf	21000000012
Deverman_Area Based Quantification of Editing Efficiency Protocol	Procedure for non-IHC based image quantification of editing.	Deverman_Area_based_quantification_of_editing_efficiency.pdf	21000000011

Experiment Record Detail