Journal: Molecular Therapy. Nucleic Acids

Article Title: Ineffective behavioral rescue despite partial brain Dp427 restoration by AAV9-U7-mediated exon 51 skipping in mdx52 mice

doi: 10.1016/j.omtn.2025.102779

Figure Lengend Snippet: Dp427 restoration 9 weeks after bilateral ICV injection of AAV9-U7-51M (A) Schematic representation of the U7snRNA and its target site on dystrophin pre-mRNA. (B) Structure of the AAV vector encoding the U7snRNA cassette. The cassette is flanked by inverted terminal repeats (ITRs) and includes an engineered U7snRNA sequence (gray box) with an antisense region, driven by its natural U7 promoter (hatched box) and 3′ downstream elements (white box). (C) Overview of the experimental procedure. (D) Left: RT-PCR gel showing exon 51 skipping 9 weeks post-injection. SM: DNA ladder; Mouse 0: mdx52 mouse injected with U7-Scramble (negative control); Mice 1–7: mdx52 mice treated with a low dose of AAV9-U7-51M (1E11 vg); Mice 8–15: mdx52 mice treated with a high dose of AAV9-U7-51M (1E12 vg). Right: Quantification of exon 51 skipping via RT-qPCR, comparing low- and high-dose treatments ( n = 7 for low dose in orange and 8 for high dose in red). (E) Quantification of Dp427 protein restoration by western blot after AAV9-U7-51M injection at low and high doses expressed in a percentage of the WT ( n = 7 for low dose in orange and 8 for high dose in red). Statistical analysis (two-way ANOVA) did not reveal a dose-dependent effect due to high interindividual variability. (F) Combined analysis of Dp427 protein rescue levels irrespective of the injected dose expressed in a percentage of the WT ( n = 15) in the three brain structures.

Article Snippet: The resulting U7snRNA fragments were cloned between the inverted terminal repeats (ITRs) of a self-complementary AAV vector (pscAAV, Addgene plasmid #83279; gift from Mark Kay) for subsequent AAV production.

Techniques: Injection, Plasmid Preparation, Sequencing, Reverse Transcription Polymerase Chain Reaction, Negative Control, Quantitative RT-PCR, Western Blot

Journal: iScience

Article Title: Simple and highly specific targeting of resident microglia with adeno-associated virus

doi: 10.1016/j.isci.2024.110706

Figure Lengend Snippet: Increased long-term specificity and efficiency for the minimal hIBA1a promoter when packaged in scAAV (A) Schematic diagram of the experimental procedure. scAAV5 virus with the hIBA1a promoter was injected into sham mice or mice with L-NIO-induced stroke. Brains were analyzed 1 week or 4 weeks later. (B) Representative confocal images showing marker expression for the indicated conditions. Scale bars, 50 μm. (C) Quantifications showing microglia-specificity of GFP expression for the indicated conditions (mean ± SEM; n = 3 mice per group). (D) Quantifications showing microglia transduction efficiency for the indicated conditions (mean ± SEM; n = 3 mice per group).

Article Snippet: The self-complementary AAV (scAAV) vectors were based on the scAAV-CAG-GFP vector (Addgene #83279) by replacing the CAG promoter with the hIBA1 promoter.

Techniques: Virus, Injection, Marker, Expressing, Transduction

Journal: iScience

Article Title: Simple and highly specific targeting of resident microglia with adeno-associated virus

doi: 10.1016/j.isci.2024.110706

Figure Lengend Snippet:

Article Snippet: The self-complementary AAV (scAAV) vectors were based on the scAAV-CAG-GFP vector (Addgene #83279) by replacing the CAG promoter with the hIBA1 promoter.

Techniques: Recombinant, Software