research highlights Guiding New Generations SENSORS AND PROBES for Decades Harnessing fungal Researchers have developed the frst fully genetically encodable eukaryotic bioluminescent system.

ioluminescence abounds in the natural world. Many organisms express Bluciferases, which are that catalyze the conversion of luciferin molecules to oxyluciferins—a reaction that is so energetically efficient that excess is emitted as . Researchers use popular such as those from the and A photograph of P. pastoris expressing the the sea pansy (genus Renilla) as probes for bioluminescence genes from N. nambi. Reproduced molecular imaging because they give strong from Kotlobay et al. (2018) with permission from signal and do not require excitation light, the authors and PNAS as permitted under a unlike fluorescent proteins such as GFP. CC BY-NC-ND license (https://creativecommons. One drawback to using luciferases as org/licenses/by-nc-nd/4.0/legalcode). probes is that they require the addition of exogenous luciferins for signal generation. Worthington is supporting life This can hamper biological studies, especially science research and discovery, those involving living animals. Ilia Yampolsky was straightforward, as metabolic genes are from the Institute of Bioorganic Chemistry typically clustered in fungal genomes. and we have done so for over in Moscow, Russia, and a team of colleagues The pathway involves four enzymes that 70 years. sought to address this limitation. can be expressed in a range of organisms For the creation of a fully genetically to make them bioluminescent. Yampolsky Authoring technical manuals and encoded system, all of the enzymes required for notes that this development of autonomously guides both in print and digital synthesis of the the luciferin substrate must be glowing creatures is likely to be the main known, along with the identity of the implication of the research. Indeed, the formats. And, we are cited in —a feat that has not been achieved for authors showed that they were able to generate thousands of respected scientific any eukaryotic luciferases. Yampolsky, who has bioluminescent Pichia pastoris by inducing journals across the globe. studied light-emitting systems throughout his expression of the necessary genes. Karen career, became interested in less widely studied Sarkisyan, cofounder of Planta, a company luciferase systems, including those in fungi, that participated in the study, also highlights noting that “the molecular basis of fungal that the luciferin 3-hydroxyhispidin is stable Sharing Our Enzyme Expertise bioluminescence has not been solved for more and cell permeable, and that nnLuz gives than 50 years despite several attempts; we took signal comparable to that of Order your free copy of the it as a challenge.” in mammalian systems. NEW Tissue Dissociation Guide, The identification of bioluminescence- Sarkisyan, who is driven by the idea of related genes in fungi was made possible by generating glowing plants, says he was most 18th Edition. Simply go to: an earlier discovery of fungal luciferin by surprised by “the beauty and the simplicity the same team. “Solving its structure was of the fungal bioluminescent system.” He Worthington-Biochem.com/ cumbersome and we spent several years says, “We hope that this system will bring TissueDissociation/default.html working on it,” recalls Yampolsky. “Then bioluminescent technologies beyond being our colleagues in Krasnoyarsk suggested a used just as tools in scientific research—for crazy idea—to look for luciferin precursors example, to create glowing plants for interior in nonbioluminescent fungal species—and, and landscape design, or perhaps even for Come see us at EB ‘19 surprisingly, it worked.” street lighting. We also envision that the The next step was identifying technology will enable development of new Booth 1044 the luciferase gene from the interfaces for life–life and life–computer Neonothopanus nambi. This luciferase, communication based on light emission.” nnLuz, has no described homologs and represents a novel protein family. Yampolsky Rita Strack recalls that this was a major challenge, noting that they were unable to identify the gene Published online: 30 January 2019 from a bacterial screen and that “it was only a https://doi.org/10.1038/s41592-019-0311-4 year later that we identified the same gene by Research papers screening of cDNA libraries in yeasts.” After Kotlobay, A. A. et al. Genetically encodable discovery of the luciferase, identification bioluminescent system from fungi. Proc. Natl Acad. 800.445.9603 • 732.942.1660 of the full luciferin biosynthetic pathway Sci. USA 115, 12728–12732 (2018).

140 Nature Methods | VOL 16 | FEBRUARY 2019 | 139–145 | www.nature.com/naturemethods