Szerző szerinti böngészés "Ahmad, Joveria"

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    CRISPR/Cas9-Mediated Engineering of a Stable HEK 293T Cell Line Expressing the Hv1 Proton Channel
    Ahmad, Joveria; Naseem, Muhammad Umair; DE--Természettudományi és Technológiai Kar--Biológiai és Ökológiai Intézet
    A 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.