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a Upper panel, <t>ABCA4</t> size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.
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a Upper panel, <t>ABCA4</t> size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.
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a Upper panel, <t>ABCA4</t> size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.
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a Upper panel, <t>ABCA4</t> size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.
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a Upper panel, <t>ABCA4</t> size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.
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a Upper panel, ABCA4 size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.

Journal: Nature Communications

Article Title: mRNA trans-splicing dual AAV vectors for (epi)genome editing and gene therapy

doi: 10.1038/s41467-023-42386-0

Figure Lengend Snippet: a Upper panel, ABCA4 size and the split site (arrowhead). Lower panel, Dual REVeRT AAV.GL vectors expressing split ABCA4 . b Experimental design for ( c – e ) and Fig. S . c Representative immunostainings on retinal sections of mouse #1 (left) and mouse (#4) showing a strong or weak improvement in OCT and ERG measurements, respectively. Images of the right eye injected with dual REVeRT AAVs and images of the contralateral control eye injected with buffer solution are shown. Scale bar: 30 µm. Cyclic nucleotide-gated channel (Cngb1) antibody was used as a rod outer segment marker to evaluate ABCA4 localization. Some unspecific ABCA4 staining was detected across all sections above the outer segments. d Higher magnification of the area marked by a gray rectangle in ( c ). Scale bar: 10 µm. e Western blot from retinas of mouse #2 and mouse #3.

Article Snippet: The western blots were imaged, and the relative band intensities quantified using the ImageLab software (Bio-Rad, v5) or VisionWorks LS Analysis Software (for ABCA4, Analytik Jena).

Techniques: Expressing, Injection, Marker, Staining, Western Blot