The recent successful commercialization of therapies based on lentiviral vectors (LVs) and promising clinical studies have showcased the immense potential of LVs for disease treatment and patient benefit. LVs inherently hold significant advantages over other gene transfer agents. This includes their ability to transduce non-dividing cells, permanently alter the target cell genome, and provide stable, long-term transgene expression. Lentiviral vectors can confer high levels of gene transfer and transgene expression in a variety of cell types. However, the biodistribution and toxicity after intravenous administration have not been reported.
The RNA ISH assay enables single molecule detection of lentiviral 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. Apart from that, RNA ISH can be used for quantitative analysis of lentivirus distribution, allowing researchers to determine the number of infected cells and the amount of viral load within tissues. RNA ISH assay from Creative Bioarray allows visualization and quantification of adenovirus vector presence, transgene, and biomarker expression in treated tissue samples.
(1) Lentiviral vector DNA detection and quantification in intact fixed tissue
(2) Transgene detection and quantification in intact fixed tissue
(3) Quantifiable assays to measure percentage of cells positive for vector, transgene and biomarker expression
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