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- W2041014766 abstract "In order to increase the optical signal of genetically encoded voltage sensors, we have utilized a transposon reaction to randomly insert the front and back halves of venus into separate subunits of a voltage gated potassium channel, Kv1.4. This split fluorescent protein approach was used with the rationale that only properly folded subunits will produce a functional, fluorescent channel at the cell surface.We tested 27 combinations for optical signals using voltage clamp fluorometry in HEK293 and NIE115 (mouse neuroblastoma) cell lines. 14 combinations show fluorescence only on the plasma membrane and achieve the goal of the split can design. The best sensitivity is -0.9% in ΔF/F for a 100 mV depolarization. The on rate during depolarization is on the scale of ms, but the off rate during repolarization is very slow, on the scale of 100 ms.One combination yielded a surprising optical signal upon depolarization in NIE115 cells. The fluorescence decreased at the edge of the cell, but increased at the cell top and bottom. This phenomenon provides a clue for us to further study the mechanism of the probe's voltage sensitivity. (Funded by NIH grant 1U24NS057631-01A1)" @default.
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- W2041014766 date "2009-02-01" @default.
- W2041014766 modified "2023-09-26" @default.
- W2041014766 title "Random Insertion of Split-can Venus into Kv1.4 Yields Voltage Sensitive Fluorescent Probes" @default.
- W2041014766 doi "https://doi.org/10.1016/j.bpj.2008.12.2049" @default.
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