Independent validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.

Validation Experiment: Independent validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.

PI: Jason D Heaney, PhD>

Description: Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery. Detection of editing in non-target tissues.

Editing Assay:

Fluorescent imaging for tdTomato+ cells
Editing efficiency calculated by counting fluorescent cells (RFP) compared to DAPI stained nuclei

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

Download: Submitted files

Original Experiment/s that are being validated:

Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CMV promoter) and self complementary sgRNA vector.

Select experimental variable to highlight records on the chart:

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Results

Independent_validation_for_Asokan_Delivery_Team:_Evolving_High_Potency_AAV_Vectors_for_Neuromuscular_Genome_Editing.
											Editing Efficiency
Record Id	Condition	Tissue	Cell Type	Sex	Editor	Model	Target Locus	Guide	Vector	Dosage	Injection Frequency	Editing Efficiency Signal	Editing Efficiency % of cells	Image
15000001297	Saline Neg control, Male	heart (TARGET)	cardiac muscle cell	Male		Ai9 mouse (BCM)					once	No red cells; No red cells;
15000001298	Saline Neg control, Female	liver (TARGET)	hepatocyte	Female		Ai9 mouse (BCM)					once	No red cells; No red cells;
15000001252	cc47-SaCas9, Male	duodenum		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001291	cc47-SaCas9, Female	uterus		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	One red cell; Two red cells; No red cells; Two red cells; No red cells;
15000001293	cc47-Cre Pos control, Male	heart (TARGET)	cardiac muscle cell	Male	Cre recombinase	Ai9 mouse (BCM)			AAVcc47-Cre	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Significant red cells; Significant red cells;
15000001294	cc47-Cre Pos control, Female	liver (TARGET)	hepatocyte	Female	Cre recombinase	Ai9 mouse (BCM)			AAVcc47-Cre	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Significant red cells;
15000001246	cc47-SaCas9, Male	liver (TARGET)	hepatocyte	Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once		19.81; 23.81; 17.35; 22.66; 19.90; 24.79;
15000001247	cc47-SaCas9, Male	heart (TARGET)	cardiac muscle cell	Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once		1.93; 3.69; 1.08; 3.85; 1.3; 2.35;
15000001248	cc47-SaCas9, Male	brain		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001249	cc47-SaCas9, Male	brown adipose tissue		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	One red cell; No red cells; No red cells; A few red cells; Significant red cells; Significant red cells;
15000001250	cc47-SaCas9, Male	colon		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001251	cc47-SaCas9, Male	diaphragm		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; A few red cells;
15000001253	cc47-SaCas9, Male	epididymis		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001254	cc47-SaCas9, Male	extensor digitorum longus		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001255	cc47-SaCas9, Male	eye		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; Two red cells; No red cells; No red cells; No red cells; No red cells;
15000001256	cc47-SaCas9, Male	gastrocnemius		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001257	cc47-SaCas9, Male	ileum		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001258	cc47-SaCas9, Male	jejunum		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; Two red cells; No red cells; No red cells;
15000001259	cc47-SaCas9, Male	kidney		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001260	cc47-SaCas9, Male	lung		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	One red cell; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001261	cc47-SaCas9, Male	pancreas		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; Two red cells; No red cells; One red cell; A few red cells;
15000001262	cc47-SaCas9, Male	gonadal fat pad		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; One red cell; No red cells; No red cells; No red cells;
15000001263	cc47-SaCas9, Male	soleus muscle		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001264	cc47-SaCas9, Male	spleen		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Two red cells; A few red cells; Two red cells; A few red cells; A few red cells; A few red cells;
15000001265	cc47-SaCas9, Male	stomach		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; One red cell;
15000001266	cc47-SaCas9, Male	subcutaneous adipose tissue		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; One red cell; No red cells; No red cells; No red cells; A few red cells;
15000001267	cc47-SaCas9, Male	testis		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; Two red cells;
15000001268	cc47-SaCas9, Male	transversus abdominis muscle		Male	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells; No red cells;
15000001269	cc47-SaCas9, Female	liver (TARGET)	hepatocyte	Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once		9.27; 11.32; 6.02; 4.42; 5.84;
15000001270	cc47-SaCas9, Female	heart (TARGET)	cardiac muscle cell	Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once		4.66; 5.86; 2.9; 5.11; 5.61;
15000001271	cc47-SaCas9, Female	brain		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001272	cc47-SaCas9, Female	brown adipose tissue		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Significant red cells; Significant red cells; Significant number of red cells; Significant number of red cells; Significant number of red cells;
15000001273	cc47-SaCas9, Female	colon		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001274	cc47-SaCas9, Female	diaphragm		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001275	cc47-SaCas9, Female	duodenum		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001276	cc47-SaCas9, Female	extensor digitorum longus		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	One red cell; No red cells; No red cells; No red cells; No red cells;
15000001277	cc47-SaCas9, Female	eye		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Two red cells; One red cell; No red cells; No red cells; No red cells;
15000001278	cc47-SaCas9, Female	gastrocnemius		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001279	cc47-SaCas9, Female	ileum		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; One red cell; No red cells; No red cells; No red cells;
15000001280	cc47-SaCas9, Female	jejunum		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001281	cc47-SaCas9, Female	kidney		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	A few red cells; A few red cells; A few red cells; A few red cells; A few red cells;
15000001282	cc47-SaCas9, Female	lung		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	One red cell; A few red cells; No red cells; No red cells; No red cells;
15000001283	cc47-SaCas9, Female	pancreas		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	A few red cells; Two red cells; One red cell; One red cell; Red cell colonies; real?;
15000001284	cc47-SaCas9, Female	gonadal fat pad		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	A few red cells; No red cells; No red cells; No red cells; No red cells;
15000001285	cc47-SaCas9, Female	soleus muscle		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; One red cell; No red cells;
15000001286	cc47-SaCas9, Female	spleen		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; A few red cells; Two red cells; One red cell; No red cells;
15000001287	cc47-SaCas9, Female	stomach		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Two red cells; One red cell; No red cells; A few red cells; A few red cells;
15000001288	cc47-SaCas9, Female	subcutaneous adipose tissue		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; Significant number of red cells; No red cells; A few red cells; No red cells;
15000001289	cc47-SaCas9, Female	transversus abdominis muscle		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	No red cells; No red cells; No red cells; No red cells; No red cells;
15000001290	cc47-SaCas9, Female	ovary		Female	SauCas9	Ai9 mouse (BCM)	Ai9/Ai14 reporter transgene	Ai9 SaCas9 Guide A; Ai9 SaCas9 Guide B	AAVcc47-SaCas9-Ai9	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Multiple red cells; Significant number of red cells; No red cells; No red cells; A few red cells;
15000001292	cc47-Cre Pos control, Male	liver (TARGET)	hepatocyte	Male	Cre recombinase	Ai9 mouse (BCM)			AAVcc47-Cre	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Significant red cells; Significant red cells;
15000001295	cc47-Cre Pos control, Female	heart (TARGET)	cardiac muscle cell	Female	Cre recombinase	Ai9 mouse (BCM)			AAVcc47-Cre	1.0x10^14 vg copies/kg (vg/kg) (2.0x10^12/20g mouse)	once	Significant red cells;
15000001296	Saline Neg control, Male	liver (TARGET)	hepatocyte	Male		Ai9 mouse (BCM)					once	No red cells; No red cells;
15000001299	Saline Neg control, Female	heart (TARGET)	cardiac muscle cell	Female		Ai9 mouse (BCM)					once	No red cells; No red cells;

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

Validation Experiment: Independent validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.

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Validation Experiment: Independent validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.

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Experiment Wide Protocols