Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: In vitro assessment of new AntagoNAT sequences (A) Schematic of AAV-CMV-eGFP-miR-155-AntagoNAT-miR-155-WPRE plasmid. (B) Transient transfection revealed 13 sequences to significantly increase Scn1a expression compared to the control cells (∗ p < 0.001). Candidates K, H, and J showed the highest increase in expression (16-fold compared to control). One-way ANOVA with multiple comparison correction using Two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli. n = 9/10 for AntagoNAT sequences, n = 6 for control (3 technical replicates each). (C) Schematic of AAV-U6-AntagoNAT-CMV-eGFP-WPRE plasmid. (D) In vitro transduction of AAV9 vectors ( n = 5/6 replicates of each group, 3 technical replicates each) using either the U6 or the CMV promoter, represented in a Boxplot. One-way ANOVA, Dunnett’s multiple comparison performed.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques: In Vitro, Plasmid Preparation, Transfection, Expressing, Control, Comparison, Transduction

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: Neonatal AAV9-AntagoNAT-H gene therapy increases survival and reduces febrile seizures in DS mice Scn1a +/− mice received either AAV9-AntagoNAT-H or PBS via i.c.v. and i.v. PBS was also administered to wild-type mice ( Scn1a +/+ ). (A) Experiment timeline. (B) Survival of Scn1a +/− DS mice treated with AAV9-AntagoNAT-H ( p = 0.04), shown in percentage of survival. Log-rank (Mantel-Cox) test. (C) Weight curves, data presented as mean, with SD error bars. Two-way ANOVA with Dunnett’s multiple comparisons test. (D) Febrile seizure temperature threshold assessment, showing a significant reduction with AAV9-AntagoNAT-H treated group ( p < 0.001). Log-rank (Mantel-Cox) test. (E) Duration of febrile seizures reduced in AAV9-AntagoNAT-H group ( p < 0.001). Man-Whitney test. (F) The Racine score classification of febrile seizures observed in the experimental mice. A significant reduction in seizure severity was observed in AAV9-AntagoNAT-H group ( p = 0.02). Fisher's exact test.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques:

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: Increase of endogenous Scn1a in the cortex of AAV9-AntagoNAT-H-treated Scn1a +/− DS mice (A) Scn1a expression in the cortex, significantly increased with AAV9-AntagoNAT-H ( p = 0.032) and Scn1a +/+ PBS ( p = 0.003) compared to PBS group. (B) Scn1a expression in the heart showed a signifcant difference between Scn1a +/+ PBS and Scn1a +/− group ( p < 0.001) only. (C) Vector copy number analysis in the cortex showed a significant increase in AAV9-AntagoNAT-H group ( p < 0.001). (D) Vector copy number in the heart only revelaed a signifcant difference between Scn1a +/+ and Scn1a +/ − PBS groups ( p < 0.001). (E) Na V 1.1 expression in the cortex. One-way ANOVA Holm-Šídák’s multiple comparisons test. For each group, n values are indicated in the figure.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques: Expressing, Plasmid Preparation

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: Increase in endogenous Scn1a after i.c.v. and i.v. AAV9-AntagoNAT-H therapy to P14 Scn1a +/− DS mice (A) Schematic diagram showing the experimental plan. (B) Survival curve for AAV9-H dose D, showing percentage of survival. (C) Representative images of GFAP (top) and CD68 (middle) staining in the brain and CD68 in the liver (bottom) of animals that received AAV9-H dose D. Further details in . (D) Survival curve of animals that received AAV9-H dose E, showing percentage of survival. Log-rank (Mantel-Cox) test. (E) Scn1a expression in cerebral cortex displayed a significant increase in both Scn1a +/+ PBS and Scn1a +/ − AAV9-AntagoNAT-H groups ( p = 0.009) compared to Scn1a +/ − PBS group. Cerebral cortex tissues for Scn1a +/− group, range from P20 to 100. (F) Na V 1.1 expression in the cerebral cortex, reported a difference between control groups only ( p = 0.013). One-way ANOVA Holm-Šídák’s multiple comparisons test.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques: Staining, Expressing, Control

Journal: Molecular Therapy Advances

Article Title: Adeno-associated virus-induced neurotoxicity is prevented by CpG depletion

doi: 10.1016/j.omta.2026.201703

Figure Lengend Snippet: CpG depletion does not alter transgene expression but prevents AAV-induced dendritic loss (A) Immunoblot of S1 homogenates probed with antibodies against transgene product EGFP and loading control GAPDH following injection of 1 × 10 10 vg of unmodified AAV1-pCAG-FLEX-EGFP or CpG-depleted AAV1-pCAG-FLEX-EGFP into Slc17a7-IRES2-Cre-D mice. Each lane contains samples from a different animal. UT, untreated. (B) Densitometry shows that EGFP protein levels are indistinguishable in ipsilateral S1 of Slc17a7-IRES2-Cre-D mice injected with either CpG-depleted or unmodified virus. EGFP protein levels are normalized to GAPDH protein levels in the same lane and represented as a fold change from the AAV1-treated condition. (C) Representative cortical neurons from untreated wild-type mice or ipsilateral (injected) hemispheres of wild-type mice injected with 1 × 10 10 vg of either unmodified or CpG-depleted AAV1-pCAG-FLEX-EGFP. Scale bars, 100 μm. (D) Total dendritic length and (E) total Sholl intersections of cortical neurons from untreated wild-type mice and wild-type mice injected with 1 × 10 10 vg of either unmodified or CpG-depleted AAV1-pCAG-FLEX-EGFP. Values are represented as a ratio of injected (ipsi) normalized to uninjected (contra) hemispheres. (F–H) Sholl intersections plotted as a function of distance from the soma for cortical neurons from the same animals shown in (E). (D and E), n = 4 untreated control mice, 10–14 neurons per animal (average = 12 cells); n = 5 mice injected with unmodified AAV1-pCAG-FLEX-EGFP, 10–14 neurons per animal (average = 12 cells); n = 5 mice injected with CpG-depleted AAV1-pCAG-FLEX-EGFP, 12–15 neurons per animal (average = 13 cells). Data in (B and D–H) are represented as mean ± SD. ∗∗∗∗ p < .0001; ∗∗∗ p < .001; ∗∗ p < .01; ns, not significant. In (G), a single ∗ indicates p < 0.05, and vertically stacked ∗ indicate p < 0.01. Statistical analysis: in (B, F–H), two-tailed unpaired t test; in (D) and (E), one-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: Male C57BL/6J and Slc17a7-IRES2-Cre-D (Strain # 037512) mice were obtained from Jackson Laboratory (The Jackson Laboratory, Bar Harbor, ME).

Techniques: Expressing, Western Blot, Control, Injection, Virus, Two Tailed Test, Comparison