CRISPR/Cas9-Mediated Engineering of a Stable HEK 293T Cell Line Expressing the Hv1 Proton Channel

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dc.contributor.advisorNaseem, Muhammad Umair
dc.contributor.authorAhmad, Joveria
dc.contributor.departmentDE--Természettudományi és Technológiai Kar--Biológiai és Ökológiai Intézet
dc.date.accessioned2025-06-16T09:17:32Z
dc.date.available2025-06-16T09:17:32Z
dc.date.created2025
dc.description.abstractA 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.
dc.description.courseBiology
dc.description.degreeMSc/MA
dc.format.extent46
dc.identifier.urihttps://hdl.handle.net/2437/391828
dc.language.isoen
dc.rights.infoHozzáférhető a 2022 decemberi felsőoktatási törvénymódosítás értelmében.
dc.subjectCRISPR/Cas9 knock-in
dc.subjectEndogenous expression
dc.subjectHEK293T
dc.subjectHv1 proton channel
dc.subjectProtein quantitation reporter
dc.subjectRPL13A locus
dc.subjectStable cell line
dc.subject.dspaceMedicine
dc.titleCRISPR/Cas9-Mediated Engineering of a Stable HEK 293T Cell Line Expressing the Hv1 Proton Channel
dc.title.subtitleEngineering of Stable HEK293T Cell Lines Expressing Hv1
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