Heterologous expression of the human potassium channel Kv2.1 in clonal mammalian cells by direct cytoplasmic microinjection of cRNA.
The cloned human delayed rectifying K+ channel Kv2.1 (drk1) was expressed in clonal mouse fibroblasts (L-cells) and rat basophilic leukemia cells (RBL-1) by direct cytoplasmic microinjection of complementary RNA (cRNA). Within six hours, cells microinjected with Kv2.1 cRNA expressed a large sustained outward current as determined from whole-cell patch-clamp recordings. Nearly 100% of cells injected with cRNA expressed outward current. Current density was 30-70 pA/pF when measured at a potential of +50 mV. Steady-state activation and inactivation parameters for Kv2.1 were similar when expressed in either L-cells or RBL-1 cells. These results are the first to demonstrate that functional ion channel proteins can be expressed in mammalian clonal cell lines by direct cytoplasmic microinjection of cRNA.
D000818:Animals; D001491:Basophils / Q000378:metabolism*; D002460:Cell Line; D003593:Cytoplasm; D004594:Electrophysiology; D005347:Fibroblasts / Q000378:metabolism*; D051379:Mice; D008845:Microinjections; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D012313:RNA / Q000008:administration & dosage / Q000235:genetics*; D018075:RNA, Complementary; D051381:Rats; D014407:Tumor Cells, Cultured
D015221:Potassium Channels; D018075:RNA, Complementary; D012313:RNA