Heterologous expression of the human potassium channel Kv2.1 in clonal mammalian cells by direct cytoplasmic microinjection of cRNA

Pflugers Arch. 1992 Nov;422(2):201-3. doi: 10.1007/BF00370422.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Basophils / metabolism*
  • Cell Line
  • Cytoplasm
  • Electrophysiology
  • Fibroblasts / metabolism*
  • Mice
  • Microinjections
  • Potassium Channels / genetics
  • Potassium Channels / metabolism*
  • Potassium Channels / physiology
  • RNA / administration & dosage
  • RNA / genetics*
  • RNA, Complementary
  • Rats
  • Tumor Cells, Cultured

Substances

  • Potassium Channels
  • RNA, Complementary
  • RNA