Mol Ther Nucleic Acids. 2020 Jun 5
Hua Yang, Keyun Qing, Geoffrey D. Keeler, Ling Yin,Mario Mietzsch, Chen Ling, Brad E. Hoffman, Mavis Agbandje-McKenna, Mengqun Tan, Wei Wang, and Arun Srivastava
Products used in the paper Details Operation
AAV vector rAAV6-CBAp-FLuc was purchased from PackGene (PackGene Biotech, Worcester, MA, USA). Request Quote

Research Field: Gene therapy/genome editing

Keywords: AAV vectors, hematopoietic stem cells, gene transfer, genome editing, hemoglobinopathies

AAV Serotype: AAV6

Dose: 1 x10^10 particles of ssAAV6-CBAp-FLuc vectors were incubated with or without PVA, in a total volume of 200 mL, at 4C for 2 h, followed by tail-vein injections (n = 5 per group). 2 weeks post-vector injections, whole-body bioluminescence imaging was performed using a Xenogen IVIS Lumina imaging system (Caliper Lifesciences, Hopkinton, MA, USA) following intraperitoneal injection of luciferin substrate (Nanolight, Pinetop, AZ, USA) at 150 mg per kg of mouse body weight. Bioluminescence images were analyzed using Living Image software (Caliper Lifesciences, Hopkinton, MA, USA). AAV6 vector genome copy numbers were determined by qPCR using total genomic DNA isolated from liver tissues.

Routes of Administration: intraperitoneal injection

Animal or cell line strain: Male C57BL/6 mice


We have reported that of the 10 most commonly used adeno-associated virus (AAV) serotype vectors, AAV6 is the most efficient in transducing primary human hematopoietic stem cells (HSCs) in vitro, as well as in vivo. More recently, polyvinyl alcohol (PVA), was reported to be a superior replacement for human serum albumin (HSA) for ex vivo expansion of HSCs. Since HSA has been shown to increase the transduction efficiency of AAV serotype vectors, we evaluated whether PVA could also enhance the transduction efficiency of AAV6 vectors in primary human HSCs. We report here that up to 12-fold enhancement in the transduction efficiency of AAV6 vectors can be achieved in primary human HSCs with PVA. We also demonstrate that the improvement in the transduction efficiency is due to PVA-mediated improved entry and intracellular trafficking of AAV6 vectors in human hematopoietic cells in vitro, as well as in murine hepatocytes in vivo. Taken together, our studies suggest that the use of PVA is an attractive strategy to further improve the efficacy of AAV6 vectors. This has important implications in the optimal use of these vectors in the potential gene therapy and genome editing for human hemoglobinopathies such as β-thalassemia and sickle cell disease.

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