Journal: Nature Communications

Article Title: The therapeutic implications of all-in-one AAV-delivered epigenome-editing platform in neurodegenerative disorders

doi: 10.1038/s41467-024-50515-6

Figure Lengend Snippet: The all-in-one AAV/ d Sa Cas9- KRAB-MeCP2(TRD) repressor platform and the control vector AAV/d Sa Cas9 with no repressor were administered by stereotaxic injection into the mouse dorsal hippocampus (DH) and validated using a GFP reporter gene ( a – c ). a , b LV-GFP reporter vector was co-injected with the AAV/gRNA1-d Sa Cas9- KRAB-MeCP2(TRD) into the left DH and with the control AAV/d Sa Cas9 into the right DH. The AAV/gRNA1-d Sa Cas9- KRAB-MeCP2(TRD) vector repressed the expression of the GFP reporter gene. a Representative images of brain coronal slices at 2x magnification 14 days and 42 days post-injection, showing GFP expression and DAPI staining in the DH. b Signals were quantified using ImageJ. Box plot displays the ratios of the left DH relative to the right DH in both age groups of 16 weeks (14d post-injection n = 9, p = 0.003; 42d post-injection n = 11, p = 0.028) and 32 weeks ( n = 10, p = 0.026) mice. Each open circle represents the quantified signal left/right for a mouse. c The AAV/gRNA1-d Sa Cas9- KRAB-MeCP2(TRD) vector repressed the expression of the GFP mRNA, box plot displays mean relative expression of GFP mRNA at 14 days ( n = 5, p = 0.027) or 42 days ( n = 6, p = 0.00046) post-injection. Each open circle represents the relative expression (log2) for a mouse. Values represent mean ± SEM. * p < 0.05 ** p < 0.01 *** p < 0.001; Two-tailed paired t -test. Source data are provided as a Source Data file.

Article Snippet: The Cj Cas9 was derived from the plasmid pX551-CMV- Cj Cas9 (addgene, #107035; gift from Alex Hewitt’s lab).

Techniques: Control, Plasmid Preparation, Injection, Expressing, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: The therapeutic implications of all-in-one AAV-delivered epigenome-editing platform in neurodegenerative disorders

doi: 10.1038/s41467-024-50515-6

Figure Lengend Snippet: The all-in-one AAV/ d Sa Cas9- KRAB-MeCP2(TRD) repressor platform and the control vector AAV/d Sa Cas9 with no repressor were administered by stereotaxic injection into the mouse dorsal hippocampus (DH) and validated using the mouse endogenous Apoe gene ( a , b ) AAV/gRNA( Apoe )p-d Sa Cas9-KRAB-MeCP2(TRD) vectors with gRNA1 or gRNA2 were injected into the right DH and the control AAV/d Sa Cas9 into the left DH. Both AAV/gRNA ( Apoe )p-d Sa Cas9-KRAB-MeCP2(TRD) vectors reduced the mouse endogenous ApoE expression. a Representative images of brain coronal slices at 2× magnification 42 days post-injection, showing ApoE expression and DAPI staining in the DH; 20× magnification of DH region showing ApoE expression. b Signals were quantified using ImageJ. Box plot displays the ratios of the right DH relative to the left DH for mice injected with the repressor vector harboring gRNA1 ( n = 8, p = 0.0002) and gRNA2 ( n = 8, p = 0.0011). Each open circle represents the quantified signal right/left for a mouse. Values represent mean ± SEM. * p < 0.05 ** p < 0.01 *** p < 0.001; Two -tailed Mann–Whitney U -Test. Source data are provided as a Source Data file.

Article Snippet: The Cj Cas9 was derived from the plasmid pX551-CMV- Cj Cas9 (addgene, #107035; gift from Alex Hewitt’s lab).

Techniques: Control, Plasmid Preparation, Injection, Expressing, Staining, Two Tailed Test, MANN-WHITNEY