Journal article

Publication year: 2021

Journal: Cells

Volume number: 10

Issue number: 5

eISSN: 2073-4409

Open access status: Gold

DOI Link: https://doi.org/10.3390/cells10051259

Publisher: MDPI

Abstract: 
Recombinant adeno-associated viruses (AAV) have emerged as an important tool for gene therapy for human diseases. A prerequisite for clinical approval is an in vitro potency assay that can measure the transduction efficiency of each virus lot produced. The AAV serotypes are typical for gene therapy bind to different cell surface structures. The binding of AAV9 on the surface is mediated by terminal galactose residues present in the asparagine-linked carbohydrates in glycoproteins. However, such terminal galactose residues are rare in cultured cells. They are masked by sialic acid residues, which is an obstacle for the infection of many cell lines with AAV9 and the respective potency assays. The sialic acid residues can be removed by enzymatic digestion or chemical treatment. Still, such treatments are not practical for AAV9 potency assays since they may be difficult to standardize. In this study, we generated human cell lines (HEK293T and HeLa) that become permissive for AAV9 transduction after a knockout of the CMP-sialic acid transporter SLC35A1. Using the human aspartylglucosaminidase (AGA) gene, we show that these cell lines can be used as a model system for establishing potency assays for AAV9-based gene therapy approaches for human diseases.

Harvard Citation style: Banning, A., Zakrzewicz, A., Chen, X., Gray, S. and Tikkanen, R. (2021) Knockout of the CMP-sialic acid transporter SLC35A1 in human cell lines increases transduction efficiency of adeno-associated virus 9: implications for gene therapy potency assays, Cells, 10(5), Article 1259. https://doi.org/10.3390/cells10051259

APA Citation style: Banning, A., Zakrzewicz, A., Chen, X., Gray, S., & Tikkanen, R. (2021). Knockout of the CMP-sialic acid transporter SLC35A1 in human cell lines increases transduction efficiency of adeno-associated virus 9: implications for gene therapy potency assays. Cells. 10(5), Article 1259. https://doi.org/10.3390/cells10051259