On-target editing compared to 14 circle-seq nominated off-target sites of adenine base editor delivered by BE-eVLP vs AAV in the C57BL/6 liver

Experiment: On-target editing compared to 14 circle-seq nominated off-target sites of adenine base editor delivered by BE-eVLP vs AAV in the C57BL/6 liver

PI: Elliot L. Chaikof, MD, PhD

Description: On-target editing compared to off-target editing at 14 CIRCLE seq nominated sites in livers of an adenine base editor delivered by engineered virus-like particles (BE-eVLPs). Treated mice vs. untreated vs. AAV was assessed one week after systemic administration of BE-eVLPs or AAV-Pcsk9 to C57BL/6 mice. DNA sequencing reads containing A-T to G-C mutations within protospacer positions 4-10.

Editing Assay: adenine base editing at target Pcsk9 site in liver vs. other mouse organs (heart, kidney, lung, muscle, spleen)

Parent Project: Delivery Technologies for In Vivo Genome Editing

Other experiments in this project: 1
- Pcsk9 adenine base editor efficiency in liver and nonliver tissue

Download: Submitted files

Jump to: | Protocols

Tissues

Liver and Biliary

Liver (TARGET)

Editors

TadA-8e V106W

Delivery Systems

eVLP

Models

C57BL/6J mouse

Sex

Male

Vectors

AAV-Pcsk9

Select experimental variable to highlight records on the chart:

Scale

Linear

Logarithmic

Note: Hover over the bars to view additional information

Results

On-target_editing_compared_to_14_circle-seq_nominated_off-target_sites_of_adenine_base_editor_delivered_by_BE-eVLP_vs_AAV_in_the_C57BL/6_liver
									Editing Efficiency
Record Id	Condition	Tissue	Sex	Editor	Model	Delivery	Vector	Dosage	Injection Frequency	Editing Efficiency % of total sequencing reads with AT converted to GC	Image
15000001532	OnTarget-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	66.86291; 63.96256; 63.03747; 56.94344;
15000001533	OT1 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.004589; 0.003633; 0.006825; 0.00658;
15000001534	OT2 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.00838; 0.010969; 0.012408; 0.004942;
15000001535	OT3 (A4)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.006714; 0.017392; 0.005735; 0.009846;
15000001536	OT4 (A4)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0; 0.004237; 0.007559; 0.014931;
15000001537	OT5 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.035982; 0.032552; 0; 0.018169;
15000001538	OT6 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.005068; 0.022923; 0.012919; 0.005606;
15000001539	OT7 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.008449; 0.007435; 0.014818; 0.007801;
15000001540	OT8 (A4)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.023479; 0.012232; 0.013158; 0;
15000001541	OT9 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.156188; 0.122231; 0.134774; 0.108782;
15000001542	OT10 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0; 0.008042; 0.003523; 0.012772;
15000001543	OT11 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.025041; 0.004222; 0.009606; 0.007518;
15000001544	OT12 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.023976; 0; 0; 0.005064;
15000001545	OT13 (A6)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.012986; 0; 0.01365; 0.008243;
15000001546	OT14 (A4)-BE-eVLP	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse	eVLP		7E11 eVLPs	once	0.012388; 0.01628; 0; 0;
15000001547	OnTarget-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	56.7267032; 58.9583333; 61.2446992; 55.0830526; 54.0576002;
15000001548	OT1 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.009653; 0.020566; 0.005427; 0.027122; 0.016123;
15000001549	OT2 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.009262; 0.008224; 0.0056; 0; 0.006272;
15000001550	OT3 (A4)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.008527; 0.013148; 0.019002; 0.016496; 0.004573;
15000001551	OT4 (A4)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.010685; 0.015029; 0.013048; 0.00508; 0.006899;
15000001552	OT5 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.008366; 0; 0; 0.015548; 0.01691;
15000001553	OT6 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.227064; 0.491477; 0.320283; 0.155862; 0.165728;
15000001554	OT7 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.220136; 0.407406; 0.280622; 0.122733; 0.164614;
15000001555	OT8 (A4)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.011838; 0.016724; 0; 0.024128; 0.016258;
15000001556	OT9 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.056104; 0.033796; 0.054093; 0.029069; 0.044902;
15000001557	OT10 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.110295; 0.248677; 0.132719; 0.085957; 0.050224;
15000001558	OT11 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.003416; 0.01301; 0; 0.003738; 0.009031;
15000001559	OT12 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.012327; 0.003763; 0.006744; 0.00948; 0.008607;
15000001560	OT13 (A6)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.002513; 0.00774; 0.006075; 0.00586; 0.012908;
15000001561	OT14 (A4)-AAV	liver (TARGET)	Male	TadA-8e V106W	C57BL/6J mouse		AAV-Pcsk9	1E11 vg	once	0.00605; 0; 0.01208; 0; 0.004069;
15000001562	OT1 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.016615; 0.004282; 0.005265;
15000001563	OT2 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.013691; 0; 0.006799;
15000001564	OT3 (A4)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0; 0.016005; 0.023632;
15000001565	OT4 (A4)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.017596; 0.008831; 0.019048;
15000001566	OT5 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.025072; 0.014474; 0.022653;
15000001567	OT6 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0; 0; 0.013784;
15000001568	OT7 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.034382; 0; 0.018874;
15000001569	OT8 (A4)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.018225; 0.010818; 0.013913;
15000001570	OT9 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.129487; 0.092557; 0.206524;
15000001571	OT10 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.008078; 0.008814; 0.019101;
15000001572	OT11 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.005991; 0.007428; 0;
15000001573	OT12 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.007818; 0; 0.005915;
15000001574	OT13 (A6)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0.029605; 0.015848; 0.010879;
15000001575	OT14 (A4)-Untreated	liver (TARGET)	Male		C57BL/6J mouse					0; 0.013443; 0.029452;

Title	Description	File Download	SCGE ID
Chaikof-Associated Protocol 2_Off-target editing AND Primers for sequencing analysis	This protocol describes in vivo adminstration and subsequent analysis of off-target editing in the liver.	Chaikof-Associated Protocol 2_Off-target editing AND Primers for sequencing analysis.pdf	21000000033

Off-target editing with BE-eVLP versus AAV treatment

DNA sequencing reads containing A-T to G-C

mutations within protospacer positions 4-10 for the 14 CIRCLE-

seq-nominated off-target loci from the livers of BE-eVLP-treated,

AAV-treated, and untreated mice. Data are shown as individual

data points and mean + =/- SEM for n=4 mice (BE-VLP), n=5 mice

(AAV), or n=3 mice (untreated). vg, viral genomes.

Contact SCGE

Experiment: On-target editing compared to 14 circle-seq nominated off-target sites of adenine base editor delivered by BE-eVLP vs AAV in the C57BL/6 liver

Results

Experiment Wide Protocols