Journal: Scientific reports

Article Title: Inhibitory luminopsins: genetically-encoded bioluminescent opsins for versatile, scalable, and hardware-independent optogenetic inhibition.

doi: 10.1038/srep14366

Figure Lengend Snippet: Figure 1. NpHR can be activated by both external light and luciferase-derived bioluminescence in HEK293 cells expressing iLMO2. (a) Emission spectra and (b) Total luminescence measured from transfected HEK293 cells expressing various luciferases (Nano-lantern, TagRFP-Rluc, Firefly) and iLMO2. (c) Schematic representation of the iLMO2 fusion protein. (d) Fluorescence image showing membrane- localized expression of iLMO2 in transfected HEK293 cells. Scale bar: 50 μ m (e) Average peak photocurrent responses to green lamp illumination and CTZ measured from HEK293 cells transfected with NpHR and Nano-lantern separately (NpHR + Nano-lantern), iLMO2 fusion protein, or NpHR alone (n = 5 for each group). Mean responses to CTZ were significantly different (*p < 0.05) but responses to green lamp illumination were not (p > 0.05) by one-way ANOVA with Bonferroni posthoc test. (f) Coupling efficiency (peak photocurrent from CTZ divided by peak photocurrent from lamp) of transfected HEK293 cells expressing NpHR and Nano-lantern separately (NpHR + Nano-lantern) (n = 5) and iLMO2 fusion protein (n = 5). Error bars indicate standard error of the mean.

Article Snippet: The luciferase constructs (TagRFP-Rluc8.638 was a gift from Dr. Sanjiv Gambhir; Firefly luciferase15 was a gift from Dr. Phil Sharp, Addgene plasmid #11510; Nano-lantern14 was a gift from 1 0Scientific RepoRts | 5:14366 | DOi: 10.1038/srep14366 Dr. Takeharu Nagai, Addgene plasmid # 51970) were all PCR amplified and cloned into the pcDNA3 vector for direct comparison of expression and bioluminescence.

Techniques: Luciferase, Derivative Assay, Expressing, Transfection, Fluorescence, Membrane