Naseem, Muhammad UmairAhmad, Joveria2025-06-162025-06-162025https://hdl.handle.net/2437/391828A stable HEK293T cell line expressing the human voltage-gated proton channel Hv1 was generated using CRISPR/Cas9-mediated knock-in at the RPL13A locus, a housekeeping gene known for consistent expression. A multicistronic cassette encoding Hv1, GFP, and a Zeocin resistance gene enabled both selection and fluorescence-based monitoring. Successful integration was confirmed by GFP expression in over 99% of cells and functional analysis using patch-clamp electrophysiology. The Hv1 current showed sensitivity to pH changes and inhibition by NZ58, confirming physiological channel behavior. This stable cell line overcomes limitations of transient expression systems and provides a reliable platform for long-term functional and pharmacological studies of Hv1.46enCRISPR/Cas9 knock-inEndogenous expressionHEK293THv1 proton channelProtein quantitation reporterRPL13A locusStable cell lineCRISPR/Cas9-Mediated Engineering of a Stable HEK 293T Cell Line Expressing the Hv1 Proton ChannelEngineering of Stable HEK293T Cell Lines Expressing Hv1MedicineHozzáférhető a 2022 decemberi felsőoktatási törvénymódosítás értelmében.