Characterization of endogenous Kv1.3 channel isoforms in T cells

Abstract

Voltage-dependent potassium channel Kv1.3 plays a key role on T cell activation; however, lack of reliable antibodies has prevented its accurate detection under endogenous circumstances. To overcome this limitation, we created a T cell-derived Jurkat cell line with endogenous Kv1.3 channel tagged in order to determine expression, location and changes upon activation of the native Kv1.3 channels. CRISPR-Cas9 technique was used to generate a double strand break on the gene locus of KCNA3 and a DNA-donor to insert a Flag-Myc peptide at the C-terminus directed by homologous recombination.