Characterization of Engineered Channelrhodopsin Variants with Improved Properties and Kinetics
作者： John Y. Lin , Michael Z. Lin , Paul Steinbach , Roger Y. Tsien
DOI: 10.1016/J.BPJ.2008.11.034
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摘要: Channelrhodopsin 2 (ChR2), a light-activated nonselective cationic channel from Chlamydomonas reinhardtii, has become useful tool to excite neurons into which it is transfected. The other ChR Chlamydomonas, ChR1, attracted less attention because of its proton-selective permeability. By making chimeras the transmembrane domains ChR1 and ChR2, combined with site-directed mutagenesis, we developed variant, named ChEF, that exhibits significantly inactivation during persistent light stimulation. ChEF undergoes only 33% inactivation, compared 77% for ChR2. Point mutation Ile170 Val (yielding “ChIEF”) accelerates rate closure while retaining reduced leading more consistent responses when stimulated above 25 Hz in both HEK293 cells cultured hippocampal neurons. In addition, these variants have altered spectral responses, sensitivity, selectivity. ChIEF allow precise temporal control depolarization, can induce action potential trains closely resemble natural spiking patterns.
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