Daniel L. Minor, Professor, Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Henry M. Colecraft, John C. Dalton Professor of Physiology and Cellular Biophysics & Cellular Biophysics, Columbia University College of Physicians & Surgeons, New York, NY, USA

• PrefaceDaniel L. Minor Jr. and Henry M. ColecraftMethods to engineer ion channels to probe different aspects of channel structure-function and regulation.1: Selection and validation of orthogonal tRNA / synthetase pairs for the encoding of unnatural amino acids across kingdoms. Christopher Ahern2: Ion channel engineering using protein trans-splicing. Stephan Alexander Pless3: ANAP: a versatile, fluorescent probe of ion channel gating and regulation. Michael C. Puljung4: Engineering voltage sensing phosphatase, VSP. Yasushi Okamura5: Probing Ion Channel Neighborhoods Using APEX. Steven Marx Methods to control and characterize ion channel function6: Controlling ion channel trafficking by targeted ubiquination and deubiquitination. Henry M. Colecraft 7: Production of recombinant venom peptides as tools for ion channel research. Eitan Reuveny8: Tethered Peptide Toxins for Ion Channels. Steven A. N. Goldstein9: Stretch and poke stimulation for characterizing mechanically activated ion channels. Jörg Grandl10: Optogenetic control of calcium influx in mammalian cells. YUBIN ZHOU11: A method for assessing ionic and molecular permeation in connexin hemichannels. JORGE E. CONTRERAS Methods to characterize ion channel function and expression in physiology and disease12: Choroid plexus epithelial cells as a model to study nongenomic steroid signaling and its effect on ion channel function. Polina V. Lishko13: Live-cell measurements of individual melanosome pH using the novel ratiometric indicator RpHiMEL. Elena Oancea 14: Long Read Transcript Profiling of Ion Channel Splice Isoforms. Liz Tunbridge15: Evaluating BEST1 Mutations in Pluripotent Stem Cell-derived Retinal Pigment Epithelial Cells. Tingting Yang16: High Throughput Electrophysiological Evaluation of Mutant Ion Channels. Alfred L. George Jr. 17: In vitro discovery of novel prokaryotic ion channel candidates for antiarrhythmic gene therapyNenad Bursac

Daniel L. Minor, Professor, Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Henry M. Colecraft, John C. Dalton Professor of Physiology and Cellular Biophysics & Cellular Biophysics, Columbia University College of Physicians & Surgeons, New York, NY, USA

• PrefaceDaniel L. Minor Jr. and Henry M. ColecraftMethods to engineer ion channels to probe different aspects of channel structure-function and regulation.1: Selection and validation of orthogonal tRNA / synthetase pairs for the encoding of unnatural amino acids across kingdoms. Christopher Ahern2: Ion channel engineering using protein trans-splicing. Stephan Alexander Pless3: ANAP: a versatile, fluorescent probe of ion channel gating and regulation. Michael C. Puljung4: Engineering voltage sensing phosphatase, VSP. Yasushi Okamura5: Probing Ion Channel Neighborhoods Using APEX. Steven Marx Methods to control and characterize ion channel function6: Controlling ion channel trafficking by targeted ubiquination and deubiquitination. Henry M. Colecraft 7: Production of recombinant venom peptides as tools for ion channel research. Eitan Reuveny8: Tethered Peptide Toxins for Ion Channels. Steven A. N. Goldstein9: Stretch and poke stimulation for characterizing mechanically activated ion channels. Jörg Grandl10: Optogenetic control of calcium influx in mammalian cells. YUBIN ZHOU11: A method for assessing ionic and molecular permeation in connexin hemichannels. JORGE E. CONTRERAS Methods to characterize ion channel function and expression in physiology and disease12: Choroid plexus epithelial cells as a model to study nongenomic steroid signaling and its effect on ion channel function. Polina V. Lishko13: Live-cell measurements of individual melanosome pH using the novel ratiometric indicator RpHiMEL. Elena Oancea 14: Long Read Transcript Profiling of Ion Channel Splice Isoforms. Liz Tunbridge15: Evaluating BEST1 Mutations in Pluripotent Stem Cell-derived Retinal Pigment Epithelial Cells. Tingting Yang16: High Throughput Electrophysiological Evaluation of Mutant Ion Channels. Alfred L. George Jr. 17: In vitro discovery of novel prokaryotic ion channel candidates for antiarrhythmic gene therapyNenad Bursac