aav2.nn  (ViGeneron GmbH)

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A ) Schematic of AAV2.NN backbone and AAV2.NN vectors used for intravitreal injection. ( B ) Bright-field and SLO image of mouse retina showing transduction and spread of vectors 3 weeks after AAV2.NN-CAG-GFP injection. ( C ) Representative retinal cross-sections immunolabeled for GFP after AAV2.NN-CAG-GFP injection for 4 weeks. ( D and E ) WB (D) and IF (E) showing expression of dCasMINI 1 month after intravitreal injection of EFS-dCasMINI into Pde6a nmf363 mice. (D) Representative immunoblots of total lysates and retina cross-sections probed with the antibody against HA. (E) Representative retinal cross sections of 8-week-old mice from each group immunolabeled for HA. ( F ) Quantitative PCR analysis of Pde6b ( N = 4) mRNA levels in sham retina and dCasMINI-treated retina. ( G ) Representative immunoblots of total sham retina and dCasMINI-treated retina lysates and detected with anti-Pde6b antibody. Data are means ± SEM. *** P < 0.001.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: Injection, Transduction, Immunolabeling, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A and B ) Representative OCT images of the sham retina, AAV2.NN-injected retina, and AAV2.NN-dCasMINI–injected retina in Pde6a nmf363 mice. The green circle showed the OCT scan area surrounding the optical nerve head. GCC indicated in the top red line, and ONL indicated in the bottom red line. ( C ) Statistics for ONL thickness. * P < 0.05. ( D ) DAPI staining of a longitudinal cryosection through the mouse retina. Staining of nuclear DNA demonstrates the retinal layers: PC, layer of outer and inner segments of photoreceptor cells. ( E ) Statistics for ONL thickness. ( F ) Representative image of sham and AAV2.NN-dCasMINI–injected retina staining with recoverin and PNA. Data are presented as means ± SEM; *** P < 0.001, two-tailed unpaired t test. OS, outer segment.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: Injection, Staining, Two Tailed Test

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A and B ) Experimental schematic of in vivo plasmid intravitreal injection at P7. ERG was obtained at 4 weeks after injection. (B) Left eye was sham group, and right eye was injected AAV2.NN. ( C ) Representative the scotopic 0.01, 0.1, and1 cd·s·m −2 dim-light rod-specific ERG traces sham eye and AAV2.NN–injected eye in Pde6a nmf363 mouse. ( D ) Quantification of maximum b-wave amplitude for (C). ( E ) Left: Constructs schematic of in vivo AAV-dCasMINI-sgPDE6B injection. Right: Left eye is sham group and right eye injected AAV2.NN-dCasMINI. ( F ) Representative the scotopic 0.01, 0.1, and 1 cd·s·m −2 dim-light rod-specific ERG traces C57BL/6 eye, AAV2.NN-dCasMINI injected eye, and sham eye in Pde6a nmf363 mouse. ( G and H ) Quantification of maximum a-wave amplitude (G) and b-wave amplitude (H) for (F). Data are presented as means ± SEM; ** P < 0.001, two-tailed unpaired t test. WPI, weeks post-immunization.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: In Vivo, Plasmid Preparation, Injection, Construct, Two Tailed Test

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A ) Experimental schematic of in vivo plasmid intravitreal injection at P7 and visual behaviors was assessed at 4 weeks after injection. ( B ) Experiment setup for OKR. ( C ) Number of responders and nonresponders is shown on the y axis with respective percentages. Statistical analysis was performed using Fisher’s exact test. * P < 0.05. ( D ) Experiment setup for looming induced escape behaviors. ( E ) Number of responders and nonresponders to looming is shown on the y axis with respective percentages. Statistical analysis was performed using Fisher’s exact test. ( F ) Statistics for response latency of AAV2.NN- ( N = 5) and dCasMINI-treated mice ( N = 11). Data are presented as means + SEM; *** P < 0.001, two-tailed unpaired t test.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: In Vivo, Plasmid Preparation, Injection, Two Tailed Test

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A ) Schematic of AAV2.NN backbone and AAV2.NN vectors used for intravitreal injection. ( B ) Bright-field and SLO image of mouse retina showing transduction and spread of vectors 3 weeks after AAV2.NN-CAG-GFP injection. ( C ) Representative retinal cross-sections immunolabeled for GFP after AAV2.NN-CAG-GFP injection for 4 weeks. ( D and E ) WB (D) and IF (E) showing expression of dCasMINI 1 month after intravitreal injection of EFS-dCasMINI into Pde6a nmf363 mice. (D) Representative immunoblots of total lysates and retina cross-sections probed with the antibody against HA. (E) Representative retinal cross sections of 8-week-old mice from each group immunolabeled for HA. ( F ) Quantitative PCR analysis of Pde6b ( N = 4) mRNA levels in sham retina and dCasMINI-treated retina. ( G ) Representative immunoblots of total sham retina and dCasMINI-treated retina lysates and detected with anti-Pde6b antibody. Data are means ± SEM. *** P < 0.001.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: Injection, Transduction, Immunolabeling, Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A and B ) Representative OCT images of the sham retina, AAV2.NN-injected retina, and AAV2.NN-dCasMINI–injected retina in Pde6a nmf363 mice. The green circle showed the OCT scan area surrounding the optical nerve head. GCC indicated in the top red line, and ONL indicated in the bottom red line. ( C ) Statistics for ONL thickness. * P < 0.05. ( D ) DAPI staining of a longitudinal cryosection through the mouse retina. Staining of nuclear DNA demonstrates the retinal layers: PC, layer of outer and inner segments of photoreceptor cells. ( E ) Statistics for ONL thickness. ( F ) Representative image of sham and AAV2.NN-dCasMINI–injected retina staining with recoverin and PNA. Data are presented as means ± SEM; *** P < 0.001, two-tailed unpaired t test. OS, outer segment.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: Injection, Staining, Two Tailed Test

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A and B ) Experimental schematic of in vivo plasmid intravitreal injection at P7. ERG was obtained at 4 weeks after injection. (B) Left eye was sham group, and right eye was injected AAV2.NN. ( C ) Representative the scotopic 0.01, 0.1, and1 cd·s·m −2 dim-light rod-specific ERG traces sham eye and AAV2.NN–injected eye in Pde6a nmf363 mouse. ( D ) Quantification of maximum b-wave amplitude for (C). ( E ) Left: Constructs schematic of in vivo AAV-dCasMINI-sgPDE6B injection. Right: Left eye is sham group and right eye injected AAV2.NN-dCasMINI. ( F ) Representative the scotopic 0.01, 0.1, and 1 cd·s·m −2 dim-light rod-specific ERG traces C57BL/6 eye, AAV2.NN-dCasMINI injected eye, and sham eye in Pde6a nmf363 mouse. ( G and H ) Quantification of maximum a-wave amplitude (G) and b-wave amplitude (H) for (F). Data are presented as means ± SEM; ** P < 0.001, two-tailed unpaired t test. WPI, weeks post-immunization.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: In Vivo, Plasmid Preparation, Injection, Construct, Two Tailed Test

Journal: Science Advances

Article Title: dCasMINI-mediated therapy rescues photoreceptors degeneration in a mouse model of retinitis pigmentosa

doi: 10.1126/sciadv.adn7540

Figure Lengend Snippet: ( A ) Experimental schematic of in vivo plasmid intravitreal injection at P7 and visual behaviors was assessed at 4 weeks after injection. ( B ) Experiment setup for OKR. ( C ) Number of responders and nonresponders is shown on the y axis with respective percentages. Statistical analysis was performed using Fisher’s exact test. * P < 0.05. ( D ) Experiment setup for looming induced escape behaviors. ( E ) Number of responders and nonresponders to looming is shown on the y axis with respective percentages. Statistical analysis was performed using Fisher’s exact test. ( F ) Statistics for response latency of AAV2.NN- ( N = 5) and dCasMINI-treated mice ( N = 11). Data are presented as means + SEM; *** P < 0.001, two-tailed unpaired t test.

Article Snippet: AAV2.NN-EGFP and AAV2.NN-EFS-dCasMINI-sgPDE6B were packaged by AAVnerGene (North Bethesda, MD).

Techniques: In Vivo, Plasmid Preparation, Injection, Two Tailed Test