This study demonstrates that CRISPR/Cas9 mediated knock-in is an effective method for creating a stable cell line expressing the Kv1.5 channel. This resolved many of the shortcomings from traditional transfection or viral transduction methods and practical for automated patch clamp. Functional characterization using patch-clamp electrophysiology confirmed the proper expression and activity of the channel. The stable model offers long-term expression even after cryopreservation, making it a reliable platform for future studies on the Kv1.5 channel and its pharmacological modulation.