Development of cell lines with increased susceptibility to diverse adeno-associated viral vectors to enable in vitro potency assays.

ABSTRACT: Vectors based on adeno-associated viruses (AAVs) are promising therapeutic modalities used in gene therapy. Robust cell-based assays that demonstrate and quantify the potency of AAV vectors in expressing their transgene are needed for clinical development. However, many AAV clinical serotypes poorly transduce cells in vitro and often contain cell-type-specific promoters inactive in commonly used cell lines. Here, we enhance the efficiency of in vitro AAV transduction by overexpressing the AAV receptor (AAVR/KIAA0319L), preventing transcriptional silencing by the HUSH complex, and using CRISPR activation (CRISPRa) to drive transgene expression. For the latter, we utilized guide RNAs targeting the conserved AAV2 inverted terminal repeat (ITR) sequence present in most AAV transf

SUBMITTER: Zengel J 

PROVIDER: S-EPMC11851225 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature