The use of the rat LAP LCR and promoter for the high-level constitutive expression of K+ channel cDNAs in a rat liver cell line.
Locus control regions (LCRs) are cis-acting elements that confer position-independent and copy-number-dependent expression upon associated genes in transgenic mice. Here we show the second example of the use of an LCR (the rat LAP LCR) in a stable expression vector system, used here in conjunction with the rat liver (NRLM) cell line. Non-transfected NRLM cells are electrically silent and highly suitable for patch clamp electrophysiology. We report reliable constitutive expression from two different K+ channel cDNAs; the voltage-gated rat clone Kv3.4 and the inward rectifier mouse clone Kir2.1. We further show that constitutive expression levels are stable for at least 8 weeks from initial recording.
Shelton
P A
PA
Department of Cell Physiology and Pharmacology, University of Leicester, UK.
Dart
C
C
D000818:Animals; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D008099:Liver / Q000166:cytology; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D015221:Potassium Channels / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D014157:Transcription Factors / Q000235:genetics*
D022762:CCAAT-Enhancer-Binding Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D015221:Potassium Channels; D012333:RNA, Messenger; D014157:Transcription Factors