Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice

Validation Experiment: Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice

PI: Jason D Heaney, PhD

Description: AAV5 encoding CRISPR/Cas editing machinery were delivered to the lungs of reporter mice by intratracheal instillation. After 4 weeks incubation, the mice were dissected and the lungs imaged for the presence of tdTomato fluorescence, indicating successful editing. Editing calculated by dividing the number of tdTomato+ red cells by the number of nuclei in each airway

Delivery Assays: PCR detection of AAV-delivered Cas9 and GFP by agarose gel electrophoresis

Editing Assay:

Fluorescent imaging for tdTomato+ cells
Editing efficiency is calculated by identifying airways in lungs sections by shape and DAPI-stained nuclei density. tdTomato-positive cells and total nuclei are counted in each airway. Airways are divided into small (<50000 square microns) and large (>50000 square microns)

Parent Project: BCM-Rice Resource for the Analysis of Somatic Gene Editing in Mice - Gao Validation (Gao, Guang-Ping )

Download: Submitted files

Original Experiment/s that are being validated:

Testing AAV5 for activation of tdTomato in mouse airway

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Results

Independent_validation_for_Gao_Delivery_Team:_Testing_ssAAV5_delivered_intratracheally_for_editing_activity_in_lung_epithelia_in_Ai9_mice_
											Editing Efficiency		Delivery Efficiency
Record Id	Condition	Tissue	Cell Type	Sex	Editor	Model	Target Locus	Guide	Vector	Dosage	Injection Frequency	Editing Efficiency Signal	Editing Efficiency % of cells	Delivery Efficiency Signal	Image
15000001042	ssAAV5-sgB.saCas9, Ai9 - SauSpyCas9, male	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Male	SaCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sa_sgB	ssAAV5-sgB.saCas9	1.7E11 viral genomes	once	< 10 red cells detected; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected; Sporadic red cells not concentrated in airway epithelia;
15000001043	ssAAV5-sgB.saCas9, Ai9 - SauSpyCas9, male	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Male	SaCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sa_sgB	ssAAV5-sgB.saCas9	1.7E11 viral genomes	once	< 10 red cells detected; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected; Sporadic red cells not concentrated in airway epithelia;
15000001044	ssAAV5-sgB.saCas9, Ai9 - SauSpyCas9, male	lung (TARGET)	respiratory epithelial cell	Male	SaCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sa_sgB	ssAAV5-sgB.saCas9	1.7E11 viral genomes	once			Negative; Negative; Negative; Positive (SaCas9);
15000001045	ssAAV5-sgB.saCas9, Ai9 - SauSpyCas9, female	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Female	SaCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sa_sgB	ssAAV5-sgB.saCas9	1.7E11 viral genomes	once	Sporadic red cells not concentrated in airway epithelia; Sporadic red cells not concentrated in airway epithelia; Sporadic red cells not concentrated in airway epithelia; Sporadic red cells not concentrated in airway epithelia; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected;
15000001046	ssAAV5-sgB.saCas9, Ai9 - SauSpyCas9, female	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Female	SaCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sa_sgB	ssAAV5-sgB.saCas9	1.7E11 viral genomes	once	Sporadic red cells not concentrated in airway epithelia; Sporadic red cells not concentrated in airway epithelia; Sporadic red cells not concentrated in airway epithelia; Sporadic red cells not concentrated in airway epithelia; < 10 red cells detected; < 10 red cells detected; < 10 red cells detected;
15000001047	ssAAV5-sgB.saCas9, Ai9 - SauSpyCas9, female	lung (TARGET)	respiratory epithelial cell	Female	SaCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sa_sgB	ssAAV5-sgB.saCas9	1.7E11 viral genomes	once			Positive (SaCas9); Positive (SaCas9);
15000001048	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, male	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Male	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once		21.5;
15000001049	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, male	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Male	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once		17.6;
15000001050	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, male	lung (TARGET)	respiratory epithelial cell	Male	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once			Negative; Negative; Negative; Negative; Negative; Positive (SpCas9 and GFP);
15000001051	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once		17.6; 21.2; 17.3; 18.2;
15000001052	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once		14.5; 11.6; 13.9; 14.6;
15000001053	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	pancreas		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; no red cells;
15000001054	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	ovary		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	some clusters of red cells but signal is weak; no red cells; no red cells; no red cells; no red cells;
15000001055	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	uterus		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	maybe a few weakly red cells; no red cells; no red cells; no red cells;
15000001056	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	trachea (TARGET)		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	significant red cells along tracheal lining; one red cell along tracheal lining; red cells along tracheal lining; red cells along tracheal lining;
15000001057	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	kidney		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001058	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	brown adipose tissue		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	No red cells; No red cells; No red cells; no red cells; no red cells;
15000001059	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	gonadal fat pad		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; no red cells;
15000001060	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	subcutaneous adipose tissue		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; two potential red cell clusters;
15000001061	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	brain		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; one red cell; no red cells; no red cells; no red cells;
15000001062	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	eye		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; no red cells;
15000001063	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	spleen		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	maybe one red cell; No red cells; No red cells; No red cells; No red cells;
15000001064	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	diaphragm		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001065	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	extensor digitorum longus		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	No red cells; no red cells; no red cells; no red cells; no red cells;
15000001066	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	gastrocnemius		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; a few faint red fibers; no red cells; no red cells;
15000001067	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	soleus muscle		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; no red cells;
15000001068	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	transversus abdominis muscle		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells;
15000001069	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	liver		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	one small cluster (~5) of red cells; no red cells; two red cells together; No red cells; No red cells;
15000001070	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	heart		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	No red cells; One red cell; No red cells; No red cells; No red cells;
15000001071	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	colon		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	No red cells; No red cells; No red cells; No red cells; no red cells;
15000001072	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	duodenum		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; no red cells;
15000001073	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	ileum		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; two clusters of red epithelial cells at vili tips; no red cells;
15000001074	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	jejunum		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; no red cells; no red cells;
15000001075	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	stomach		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once	no red cells; no red cells; no red cells; one red cell; small cluster of red cells;
15000001076	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9 - SauSpyCas9, female	lung (TARGET)		Female	SpCas9	Ai9-SauSpyCas9 mouse	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once			Negative; Negative; Positive (SpCas9 and GFP); Positive (SpCas9 and GFP); Positive (SpCas9 and GFP);
15000001077	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9, female	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Female	SpCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once		22.1; 25.3;
15000001078	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9, female	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Female	SpCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once		19.3; 20.6;
15000001079	ssAAV5-spCas9 + ssAAV5-sgA+sgB-U1A.GFP, Ai9, female	lung (TARGET)	respiratory epithelial cell	Female	SpCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	BCM_ssAAV5-Sp_sgB; BCM_ssAAV5-Sp_sgA	ssAAV5-sgA+sgB-U1A.GFP ; ssAAV5-spCas9	1.7E11 viral genomes	once			Positive (SpCas9 and GFP); Positive (SpCas9 and GFP);
15000001080	scAAV5-Cre-GFP Pos Control, Ai9 - SauSpyCas9, male	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Male	Cre recombinase	Ai9-SauSpyCas9 mouse			scAAV5-Cre-GFP	1.7E11 viral genomes	once	Significant red cells; Significant red cells;
15000001081	scAAV5-Cre-GFP Pos Control, Ai9 - SauSpyCas9, male	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Male	Cre recombinase	Ai9-SauSpyCas9 mouse			scAAV5-Cre-GFP	1.7E11 viral genomes	once	Significant red cells; Significant red cells;
15000001082	scAAV5-Cre-GFP Pos Control, Ai9 - SauSpyCas9, male	lung (TARGET)	respiratory epithelial cell	Male	Cre recombinase	Ai9-SauSpyCas9 mouse			scAAV5-Cre-GFP	1.7E11 viral genomes	once			Negative; Negative;
15000001083	scAAV5-Cre-GFP Pos Control, Ai9 - SauSpyCas9, female	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Female	Cre recombinase	Ai9-SauSpyCas9 mouse			scAAV5-Cre-GFP	1.7E11 viral genomes	once	Significant red cells; Significant red cells;
15000001084	scAAV5-Cre-GFP Pos Control, Ai9 - SauSpyCas9, female	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Female	Cre recombinase	Ai9-SauSpyCas9 mouse			scAAV5-Cre-GFP	1.7E11 viral genomes	once	Significant red cells; Significant red cells;
15000001085	scAAV5-Cre-GFP Pos Control, Ai9 - SauSpyCas9, female	lung (TARGET)	respiratory epithelial cell	Female	Cre recombinase	Ai9-SauSpyCas9 mouse			scAAV5-Cre-GFP	1.7E11 viral genomes	once			Negative; Negative;
15000001086	saline Neg control, male	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001087	saline Neg control, male	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001088	saline Neg control, male	pancreas		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001089	saline Neg control, male	epididymis		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001090	saline Neg control, male	testis		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001091	saline Neg control, male	trachea (TARGET)		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001092	saline Neg control, male	kidney		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; no red cells;
15000001093	saline Neg control, male	brown adipose tissue		Male		Ai9-SauSpyCas9 mouse					once	no red cells; one red cell?; no red cells;
15000001094	saline Neg control, male	gonadal fat pad		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001095	saline Neg control, male	subcutaneous adipose tissue		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001096	saline Neg control, male	brain		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001097	saline Neg control, male	eye		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001098	saline Neg control, male	spleen		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001099	saline Neg control, male	diaphragm		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001100	saline Neg control, male	extensor digitorum longus		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001101	saline Neg control, male	gastrocnemius		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; a few red cells;
15000001102	saline Neg control, male	soleus muscle		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001103	saline Neg control, male	transversus abdominis muscle		Male		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; no red cells;
15000001104	saline Neg control, male	liver		Male		Ai9-SauSpyCas9 mouse					once	Two red cell cluster; Two red cell cluster; no red cells;
15000001105	saline Neg control, male	heart		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001106	saline Neg control, male	colon		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001107	saline Neg control, male	duodenum		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001108	saline Neg control, male	ileum		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001109	saline Neg control, male	jejunum		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001110	saline Neg control, male	stomach		Male		Ai9-SauSpyCas9 mouse					once	no red cells; no red cells; no red cells;
15000001111	saline Neg control, male	lung (TARGET)		Male		Ai9-SauSpyCas9 mouse					once			Negative; Negative; Negative;
15000001112	saline Neg control, female	epithelium of bronchiole (TARGET)	respiratory epithelial cell	Female		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001113	saline Neg control, female	epithelium of main bronchus (TARGET)	respiratory epithelial cell	Female		Ai9-SauSpyCas9 mouse					once	No red cells; No red cells; No red cells;
15000001114	saline Neg control, female	pancreas		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001115	saline Neg control, female	epididymis		Female		Ai9-SauSpyCas9 mouse					once	;
15000001116	saline Neg control, female	ovary		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001117	saline Neg control, female	testis		Female		Ai9-SauSpyCas9 mouse					once	;
15000001118	saline Neg control, female	uterus		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001119	saline Neg control, female	trachea (TARGET)		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001120	saline Neg control, female	kidney		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001121	saline Neg control, female	brown adipose tissue		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001122	saline Neg control, female	gonadal fat pad		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001123	saline Neg control, female	subcutaneous adipose tissue		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001124	saline Neg control, female	brain		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001125	saline Neg control, female	eye		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001126	saline Neg control, female	spleen		Female		Ai9-SauSpyCas9 mouse					once	two red cells?;
15000001127	saline Neg control, female	diaphragm		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001128	saline Neg control, female	extensor digitorum longus		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001129	saline Neg control, female	gastrocnemius		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001130	saline Neg control, female	soleus muscle		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001131	saline Neg control, female	transversus abdominis muscle		Female		Ai9-SauSpyCas9 mouse					once	one red cell?;
15000001132	saline Neg control, female	liver		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001133	saline Neg control, female	heart		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001134	saline Neg control, female	colon		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001135	saline Neg control, female	duodenum		Female		Ai9-SauSpyCas9 mouse					once	no red cells;
15000001136	saline Neg control, female	ileum		Female		Ai9-SauSpyCas9 mouse					once	No red cells;
15000001137	saline Neg control, female	jejunum		Female		Ai9-SauSpyCas9 mouse					once	No red cells;
15000001138	saline Neg control, female	stomach		Female		Ai9-SauSpyCas9 mouse					once	No red cells;
15000001139	saline Neg control, female	lung (TARGET)		Female		Ai9-SauSpyCas9 mouse					once			Negative; Negative; Negative;

Title	Description	File Download	SCGE ID
Heaney_SATC Tissue Processing, Imaging and Analysis	Procedure for tissue preparation, imaging and analysis.	Heaney_SATC_Tissue_Processing_Imaging_and_Analysis.pdf	21000000025
Heaney-SATC_Gao-Validation_Intratracheal Delivery of AAV in Mice	Procedure for Intratracheal (IT) delivery of AAV in mouse lung.	Heaney-SATC_Gao-Validation_Intratracheal_Delivery_of_AAV_in_Mice.pdf	21000000023

Validation Experiment: Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice

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Validation Experiment: Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice

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Results

Experiment Wide Protocols