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Experiment: Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.

PI:   Aravind Asokan, PhD 

Description:  C57BL/6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding a GFP reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
Delivery Assays:
  • Native fluorescence was quantified using Image J. For each animal, 6 images were taken from 2 sections and used to calculate corrected total cell fluorescence using the formula: Integrated Density – (Area of selected image X Mean fluorescence of background readings).
  • Relative neuronal transduction levels were quantified by counting the number of transduced neurons, identified based on morphology, for each brain region per 50um sagittal section
Parent Project: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
Download: Submitted files
Filter Results ..
  Tissues
Renal/Urinary
  • Kidney
Liver and Biliary
  • Liver
Cardiovascular
  • Heart
Musculoskeletal
  • Skeletal muscle tissue
Nervous
  • Cerebral cortex (neuron)
  • Cerebral cortex (astrocyte)
  • Cerebellum (neuron)
  • Cerebellum (astrocyte)
  • Hippocampal formation (neuron)
  • Hippocampal formation (astrocyte)
  • Dorsal plus ventral thalamus (neuron)
  • Dorsal plus ventral thalamus (astrocyte)
  • Striatum (neuron)
  • Striatum (astrocyte)
  • Midbrain (neuron)
  • Midbrain (astrocyte)
  Cell Types
 astrocyte
 neuron
Models
 C57BL/6 mouse (Asokan study)
Sex
 Male
  Vectors
 AAV9-GFP
 AAVcc81-GFP
 AAVcc84-GFP
Select experimental variable to highlight records on the chart:

Note: Hover over the bars to view additional information

Results
Testing_virus_region_8_(VR8)_mutant_cross-species_compatible_Adeno_Associated_Viruses_(ccAAVs)_in_mice._
Delivery Efficiency
Record Id Condition Tissue Cell Type Sex Model Vector Dosage Injection Frequency Delivery Efficiency # of cells in 50um Section Delivery Efficiency Corrected Total Fluorescence Image
15000000363 AAV9 GFP cerebral cortex neuron Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000364 AAV9 GFP cerebral cortex astrocyte Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000365 AAV9 GFP cerebellum neuron Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000366 AAV9 GFP cerebellum astrocyte Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000367 AAV9 GFP hippocampal formation neuron Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000368 AAV9 GFP hippocampal formation astrocyte Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000369 AAV9 GFP dorsal plus ventral thalamus neuron Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000370 AAV9 GFP dorsal plus ventral thalamus astrocyte Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000371 AAV9 GFP striatum neuron Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000372 AAV9 GFP striatum astrocyte Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000373 AAV9 GFP midbrain neuron Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000374 AAV9 GFP midbrain astrocyte Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000375 AAV9 GFP kidney Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000376 AAV9 GFP liver Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000377 AAV9 GFP heart Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000378 AAV9 GFP skeletal muscle tissue Male C57BL/6 mouse (Asokan study) AAV9-GFP 5e13 vg/kg 1X
15000000379 AAVcc84 cerebral cortex neuron Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000380 AAVcc84 cerebral cortex astrocyte Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000381 AAVcc84 cerebellum neuron Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000382 AAVcc84 cerebellum astrocyte Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000383 AAVcc84 hippocampal formation neuron Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000384 AAVcc84 hippocampal formation astrocyte Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000385 AAVcc84 dorsal plus ventral thalamus neuron Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000386 AAVcc84 dorsal plus ventral thalamus astrocyte Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000387 AAVcc84 striatum neuron Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000388 AAVcc84 striatum astrocyte Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000389 AAVcc84 midbrain neuron Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000390 AAVcc84 midbrain astrocyte Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000391 AAVcc84 liver Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000392 AAVcc84 heart Male C57BL/6 mouse (Asokan study) AAVcc84-GFP 5e13 vg/kg 1X
15000000393 AAVcc81 cerebral cortex neuron Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000394 AAVcc81 cerebral cortex astrocyte Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000395 AAVcc81 cerebellum neuron Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000396 AAVcc81 cerebellum astrocyte Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000397 AAVcc81 hippocampal formation neuron Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000398 AAVcc81 hippocampal formation astrocyte Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000399 AAVcc81 dorsal plus ventral thalamus neuron Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000400 AAVcc81 dorsal plus ventral thalamus astrocyte Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000401 AAVcc81 striatum neuron Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000402 AAVcc81 striatum astrocyte Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000403 AAVcc81 midbrain neuron Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000404 AAVcc81 midbrain astrocyte Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000405 AAVcc81 kidney Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000406 AAVcc81 liver Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000407 AAVcc81 heart Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
15000000408 AAVcc81 skeletal muscle tissue Male C57BL/6 mouse (Asokan study) AAVcc81-GFP 5e13 vg/kg 1X
Expression of VR8 ccAAVs in the brain following IV injection

Expression of VR8 ccAAVs in the brain. Adult C57/B6 mice were injected intravenously at 5e13 vg/kg (N=3). A self-complementary GFP cassette was packaged into lead ccAAV mutants driven by a chicken-beta actin human hybrid (CBh) promoter. Mice were sacrificed 4-weeks post injection brain was post-fixed in 4% PFA for 24 hrs. Tissue was cut into 50mm sagittal sections using a vibratome. Immunohistochemical staining of brain sections revealed neuronal and astrocytic expression (representative images are shown)
Expression of VR8 ccAAVs in cardiac and skeletal muscle

Expression of VR8 ccAAVs in cardiac and skeletal muscle. Adult C57/B6 mice were injected intravenously at 5e13 vg/kg (N=3). A self-complementary GFP cassette was packaged into lead ccAAV mutants driven by a chicken-beta actin human hybrid (CBh) promoter. Mice were sacrificed 4-weeks post injection and heart and skeletal muscle was post-fixed in 4% PFA for 24 hrs. Fixed tissue was embedded in agarose and cut into 50mm thick sections using a vibratome. Native fluorescence was quantified using Image J. For each animal, 6 images were taken from 2 sections and used to calculate corrected total cell fluorescence using the formula: Integrated Density (Area of selected image X Mean fluorescence of background readings).
Expression of VR8 ccAAVs in liver and kidney

Expression of VR8 ccAAVs in liver and kidney. Adult C57/B6 mice were injected intravenously at 5e13 vg/kg (N=3). A self-complementary GFP cassette was packaged into lead ccAAV mutants driven by a chicken-beta actin human hybrid (CBh) promoter. Mice were sacrificed 4-weeks post injection and liver and kidney was post-fixed in 4% PFA for 24 hrs. Fixed tissue was embedded in agarose and cut into 50mm thick sections using a vibratome. Native fluorescence was quantified using Image J. For each animal, 6 images were taken from 2 sections and used to calculate corrected total cell fluorescence using the formula: Integrated Density (Area of selected image X Mean fluorescence of background readings).