4 files

jASAP voltage indicator optimization

posted on 10.05.2022, 19:51 authored by Jeremy HassemanJeremy Hasseman, Getahun Tsegaye, Daniel Reep, Arthur Tsang, Ilya KolbIlya Kolb, Allan Wong, Amy Chuong, Jihong ZhengJihong Zheng, Ben ArthurBen Arthur, Wyatt KorffWyatt Korff, GENIE Project Team

ASAPs (Accelerated Sensor of Action Potentials), developed by Michael Lin (https://doi.org/10.1038/nn.3709, https://doi.org/10.1016/j.cell.2019.11.004), represents a class of voltage indicators that’s sensitive (from spike detection to subthreshold membrane potential reporting), fast (submillisecond level for recently published ASAP2f variant) and two-photon compatible. GENIE project team has put effort in optimizing ASAP voltage indicator. With our high-throughput HEK cell screening  platform we identified one top hit, T392V mutation based on ASAP3,  dramatically improve the sensitivity (by ~60%) without compromising  kinetics. ASAP (T392V) variants performance was further tested in mammalian neurons (field stimulation screening and voltage clamp assay) and mouse visual cortex in vivo.