Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain

Validation Experiment: Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain

PI: Stephen A Murray, PhD

Description: Delivery of CRISPR/Cas9 via ribonuclear protein (RNP) loaded nanocages (NC) to the brain in Ai14 mice by intracranial bilateral injection. Tissues were harvested 14 days after NC administeration. On-target and off-target editing was assessed.

Editing Assay: On-target editing efficiency was calculated by counting the number of edited Neurons (NeuN+/GFP+ and tdTomato+/RFP+) out of the total number of neurons (NeuN+/GFP+) in a defined region of interest in the brain, expressed as a percentage of neurons in the region of interest (ROI). Off-target editing analysis: peripheral tissues were fixed, cryosectioned and imaged directly for RFP (tdTomato). Presence or absence of signal above background was recorded for each tissue assessed. Biological replicates by condition were as follows: NC-CPP Males: On-target N=3, Off-target N=1; NC-CPP Females: On-target N=3, Off-target N=1; NC-RVG Males: On-target N=3, Off-target N=1; NC-RVG Females: On-target N=3, Off-target N=2; NC No Ligand Male: On-target N=3, Off-target N=2; NC No Ligand Female: On-target N=3, Off-target N=1. *Inconclusive: Signal in cells surrounding epididymal duct/testis germinal epithelium was observed in one animal (1 out of 3-4 tissue sections)

Parent Project: The Jackson Laboratory Gene Editing Testing Center (JAX-GETC) - SATC-Gong, Submission 2 (On- and Off-target data)

Download: Submitted files

Original Experiment/s that are being validated:

Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.

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Results

Independent_validation_of_Gong_delivery_platform_using_RNP-loaded_nanocages_to_deliver_CRISPR/Cas9_to_mouse_brain
										Editing Efficiency
Record Id	Condition	Tissue	Cell Type	Sex	Editor	Model	Delivery	Target Locus	Guide	Injection Frequency	Editing Efficiency % cells	Editing Efficiency Signal	Image
15000003294	NC-CPP Males	striatum (TARGET)	neuron	Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once	25.7; 28.5; 24.7;
15000003295	NC-CPP Males	urinary bladder		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003296	NC-CPP Males	eye		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003297	NC-CPP Males	gastrocnemius		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003298	NC-CPP Males	heart		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003299	NC-CPP Males	kidney		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003300	NC-CPP Males	large intestine		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003301	NC-CPP Males	liver		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003302	NC-CPP Males	lung		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003303	NC-CPP Males	pancreas		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003304	NC-CPP Males	skin of body		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003305	NC-CPP Males	small intestine		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003306	NC-CPP Males	spleen		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003307	NC-CPP Males	stomach		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003308	NC-CPP Males	thymus		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003309	NC-CPP Males	epididymis		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003310	NC-CPP Males	testis		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003311	NC-CPP Females	striatum (TARGET)	neuron	Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once	26.1; 26.4; 25.6;
15000003312	NC-CPP Females	urinary bladder		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003313	NC-CPP Females	eye		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003314	NC-CPP Females	gastrocnemius		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003315	NC-CPP Females	heart		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003316	NC-CPP Females	kidney		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003317	NC-CPP Females	large intestine		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003318	NC-CPP Females	liver		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003319	NC-CPP Females	lung		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003320	NC-CPP Females	pancreas		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003321	NC-CPP Females	small intestine		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003322	NC-CPP Females	spleen		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003323	NC-CPP Females	stomach		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003324	NC-CPP Females	thymus		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003325	NC-CPP Females	uterus		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003326	NC-CPP Females	ovary		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-CPP	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003327	NC-RVG Males	striatum (TARGET)	neuron	Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once	20.4; 24.4; 21.7;
15000003328	NC-RVG Males	urinary bladder		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003329	NC-RVG Males	eye		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003330	NC-RVG Males	gastrocnemius		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003331	NC-RVG Males	heart		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003332	NC-RVG Males	kidney		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003333	NC-RVG Males	large intestine		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003334	NC-RVG Males	liver		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003335	NC-RVG Males	lung		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003336	NC-RVG Males	pancreas		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003337	NC-RVG Males	skin of body		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003338	NC-RVG Males	small intestine		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003339	NC-RVG Males	spleen		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003340	NC-RVG Males	stomach		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003341	NC-RVG Males	thymus		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003342	NC-RVG Males	epididymis		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		inconclusive*;
15000003343	NC-RVG Males	testis		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003344	NC-RVG Females	striatum (TARGET)	neuron	Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once	30.2; 27.8; 24.5;
15000003345	NC-RVG Females	urinary bladder		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003346	NC-RVG Females	eye		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003347	NC-RVG Females	gastrocnemius		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003348	NC-RVG Females	heart		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003349	NC-RVG Females	kidney		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003350	NC-RVG Females	large intestine		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003351	NC-RVG Females	liver		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003352	NC-RVG Females	lung		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003353	NC-RVG Females	pancreas		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003354	NC-RVG Females	skin of body		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003355	NC-RVG Females	small intestine		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003356	NC-RVG Females	spleen		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003357	NC-RVG Females	stomach		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003358	NC-RVG Females	thymus		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003359	NC-RVG Females	uterus		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003360	NC-RVG Females	ovary		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-RVG	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003361	NC No Ligand Male	striatum (TARGET)	neuron	Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once	16.7; 30.4; 18.9;
15000003362	NC No Ligand Male	urinary bladder		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003363	NC No Ligand Male	eye		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003364	NC No Ligand Male	gastrocnemius		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003365	NC No Ligand Male	heart		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003366	NC No Ligand Male	kidney		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003367	NC No Ligand Male	large intestine		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003368	NC No Ligand Male	liver		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003369	NC No Ligand Male	lung		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003370	NC No Ligand Male	pancreas		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003371	NC No Ligand Male	skin of body		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003372	NC No Ligand Male	small intestine		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003373	NC No Ligand Male	spleen		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003374	NC No Ligand Male	stomach		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003375	NC No Ligand Male	thymus		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; absent;
15000003376	NC No Ligand Male	epididymis		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003377	NC No Ligand Male	testis		Male	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent; inconclusive*;
15000003378	NC No Ligand Female	striatum (TARGET)	neuron	Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once	25.0; 27.6; 34.0;
15000003379	NC No Ligand Female	urinary bladder		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003380	NC No Ligand Female	eye		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003381	NC No Ligand Female	gastrocnemius		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003382	NC No Ligand Female	heart		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003383	NC No Ligand Female	kidney		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003384	NC No Ligand Female	large intestine		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003385	NC No Ligand Female	liver		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003386	NC No Ligand Female	lung		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003387	NC No Ligand Female	pancreas		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003388	NC No Ligand Female	skin of body		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003389	NC No Ligand Female	small intestine		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003390	NC No Ligand Female	spleen		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003391	NC No Ligand Female	stomach		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003392	NC No Ligand Female	thymus		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003393	NC No Ligand Female	uterus		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;
15000003394	NC No Ligand Female	ovary		Female	sNLS-SpCas9-sNLS	Ai14 mouse (congenic)	RNP-NC-no ligand	Ai9/Ai14 reporter transgene	Ai14 gRNA	once		absent;

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Validation Experiment: Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain

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