Automated determination of 8-OHdG in cells and tissue via immunofluorescence using a specially created antibody

Jung, Tobias; Findik, Nicole; Hartmann, Bianca; Hanack, Katja; Grossmann, Kai; Roggenbuck, Dirk; Wegmann, Marc; Mantke, René; Deckert, Markus; Grune, Tilman

doi:10.1016/j.btre.2024.e00833

Biotechnology Reports

2024

DOI: 10.1016/j.btre.2024.e00833

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Automated determination of 8-OHdG in cells and tissue via immunofluorescence using a specially created antibody

Tobias Jung,

Nicole Findik,

Bianca Hartmann

et al.

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2024

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Cited by 2 publications

References 66 publications

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Promising tools into oxidative stress: A review of non-rodent model organisms

Zhang,

Li,

Ren

et al. 2024

Redox Biology

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Promising tools into oxidative stress: A review of non-rodent model organisms

Zhang,

Li,

Ren

et al. 2024

Redox Biology

View full text Add to dashboard Cite

Retinal Penetrating Adeno-Associated Virus

Kumar,

Mishra,

Cashman

et al. 2024

Invest. Ophthalmol. Vis. Sci.

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Purpose The most common method of delivery of genes to the outer retina uses recombinant adeno-associated virus (AAV) injected into the subretinal space using a surgical procedure. In contrast, most drugs are delivered to the retina using an intravitreal approach in an office setting. The objective of the current study was to develop AAV vectors that can reach the outer retina via intravitreal injection. Methods Recently, we described a molecular chaperone (Nuc1) that enhanced the penetration of small and large molecules, including AAV, into the retina. The Nuc1 amino acid sequence or a truncated version of Nuc1 (IKV) was genetically incorporated into an exposed loop of AAV2/9 VP1 protein. These novel recombinant AAV vectors expressing green fluorescent protein (GFP) or nuclear factor erythroid 2 p45-related factor 2 (Nrf2) were injected into the vitreous of C57Bl/6J or Nrf2 knockout mice, respectively. The amount of GFP expression or oxidative stress as measured by 8-Hydroxy-2′-deoxyguanosine staining in C57Bl/6J or Nrf2 knockout mice, respectively, was quantified. Results Incorporation of Nuc1 into AAV2/9 did not lead to significant expression of GFP in the murine retina. However, incorporation of IKV into AAV2/9 led to robust expression of GFP in photoreceptors and retinal pigment epithelium (RPE) via the intravitreal and subretinal routes of delivery. Furthermore, expression of Nrf2 using an IKV vector led to a reduction in oxidative stress in the retina of C57Bl/6J and Nrf2 knockout mice. Conclusions We have developed a novel AAV vector that enables delivery of transgenes to the outer retina of mice, including photoreceptors and RPE following intravitreal injection.

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Automated determination of 8-OHdG in cells and tissue via immunofluorescence using a specially created antibody

Cited by 2 publications

References 66 publications

Promising tools into oxidative stress: A review of non-rodent model organisms

Promising tools into oxidative stress: A review of non-rodent model organisms

Retinal Penetrating Adeno-Associated Virus

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