Adeno-associated virus (AAV)-based vectors have proven to be effective vehicles for efficient and stable gene delivery with broad tropism in vivo. Determining the AAV vector distribution is central for the safety assessment of proposed early-phase clinical trials. The RNA ISH assay enables single molecule detection of AAV vector DNA, transgene mRNA, and/or specific biomarkers in the tissue at the single-cell level, thus allowing for a high spatial resolution at both the cellular and subcellular levels. Cell type-specific promoter activity, transgene, and cell-specific biomarker expression heterogeneity can be assessed with the RNA ISH assays, as can screening of vector delivery systems for cellular uptake efficiency and extracellular trapping of the vector. RNA ISH assay from Creative Bioarray allows visualization and quantification of AAV vector presence, transgene, and biomarker expression in treated tissue samples.
We are capable of offering AAV Vector/Transgene/Biomarker Quantification and Statistical Analysis service in the most high-quality and cost-effective way. Following the current FDA guidelines, our experienced scientists will design the most appropriate experiment plan to meet every client's specific requirement.
Figure 1. AAV-treated liver stained with the RNAscope 2.5 LS Red assay. (A) AAV vector primarily sequestered in the interstitial space. (B) HALO (Indica Labs) overlay identifies AAV-positive nuclei in red and AAV-negative nuclei in white. (C) Quantification of the percentage of signal detected within the nuclear boundaries (stained with hematoxylin) vs signal located outside of the nuclear boundaries.
(1) AAV vector DNA detection and quantification in intact fixed tissue
(2) Quantifiable assays to measure percentage of cells positive for vector, transgene and biomarker expression
(3) Quantify AAV+ cell number and track persistence over time
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