Molecular biology: generation of transgenic mice, RT-PCR, cloning, site-directed mutagenesis, transcript and protein expression, immunohistochemistry and in situ hybridization, microarray analysis of gene expression.
Electrophysiology: macroscopic currents, action potentials, single and multi-unit extracellular recordings, planar multi-electrode arrays, whole-cell recording, patch-clamp, acute brain slices, dissociated primary neuronal cultures, and organotypic cultures.
Transgenic Mouse Lines
Global BK channel knockout (Slo–/–, Kcnma1–/–)
Available from Jax Labs (Stock #035902 B6.129(FVB)-Kcnma1tm1Rwa/J)
Citation: JBC 279:36746-36752 (2004)
PCR genotyping protocol & strain info
Transnetyx genotyping (Probes: Kcnma1 WT and Neomycin)
Conditional BK channel knockout, expressing tdTomato (Kcnma1fl)
Available from Jax Labs (Stock #035901 B6.Cg-Kcnma1tm1.1Alme/J)
Citation: Physiological Reports 3(11): e12612 (2015). Genbank Accession #
Transnetyx genotyping (Probes: Kcnma1-3 WT and tdRFP)
Beta2 subunit knockout, conditional, (Kcnmb2–/–), made by Chris Lingle (Wash U)
Citation: Nature Communications Mar 4;7:10837 (2016).
Original Citation for generation of mouse line: J Gen Physiol 144(4):275-95 (2014).
Available from Chris Lingle or Jax Labs (Stock #028416).
Transnetyx genotyping (Probes: Kcnmb2-1 WT and Kcnmb2-1 EX)
Genbank Links for BK Variants (Organized by Publication)
HUMAN BK CHANNEL cDNA sequences
BKVYR (MG279689) was used in the following publications:
Comparative Gain-of-Function Effects of KCNMA1-N999S Mutation on Human BK Channel Properties. J Neurophysiology 123:560 (2020).
Effects of single nucleotide polymorphisms in human KCNMA1 on BK current properties. Frontiers in Molecular Neuroscience. 12:285(2019).
MOUSE BK CHANNEL cDNA sequences
BK channels regulate sinoatrial node firing rate and cardiac pacing in vivo. AJP- Heart 307(9):H1327-38 (2014).
Phosphorylation of a constitutive serine inhibits BK channel variants containing the alternate exon ‘SRKR’. J General Physiology 142 (6):585-598 (2013).
– BKSRKR (JX462785); Mutations: BKSRKR-S642A, , BKSRKR-R640Q/R645Q
– BKSRKR with YFP tag (JX462786)
– mbr5:YFP (KF530038)
– C2:YFP (KF530044)
– C3:YFP (KF530036)
– C4:YFP, also called BKVYR (KF530042)
– BK0:YFP (KF530040); Mutations: BK0-S642D
Mis-expression of the BK K+ channel disrupts suprachiasmatic nucleus circuit rhythmicity and alters clock-controlled behavior. AJP- Cell Physiology 304(4):C299-C311 (2013).