Evaluating Brightness and Spectral Properties of Click Beetle and Firefly Luciferases Using Luciferin Analogues: Identification of Preferred Pairings of Luciferase and Substrate for In Vivo Bioluminescence Imaging. | LitMetric
Purpose: Currently, a variety of red and green beetle luciferase variants are available for bioluminescence imaging (BLI). In addition, new luciferin analogues providing longer wavelength luminescence have been developed that show promise for improved deep tissue imaging. However, a detailed assessment of these analogues (e.g., Akalumine-HCl, CycLuc1, and amino naphthyl luciferin (NH-NpLH2)) combined with state of the art luciferases has not been performed. The aim of this study was to evaluate for the first time the in vivo brightness and spectral characteristics of firefly (Luc2), click beetle green (CBG99), click beetle red 2 (CBR2), and Akaluc luciferases when paired with different D-luciferin (D-LH2) analogues in vivo.
Procedures: Transduced human embryonic kidney (HEK 293T) cells expressing individual luciferases were analyzed both in vitro and in mice (via subcutaneous injection). Following introduction of the luciferins to cells or animals, the resulting bioluminescence signal and photon emission spectrum were acquired using a sensitive charge-coupled device (CCD) camera equipped with a series of band pass filters and spectral unmixing software.
Results: Our in vivo analysis resulted in four primary findings: (1) the best substrate for Luc2, CBG99, and CBR2 in terms of signal strength was D-luciferin; (2) the spectra for Luc2 and CBR2 were shifted to a longer wavelength when Akalumine-HCl was the substrate; (3) CBR2 gave the brightest signal with the near-infrared substrate, NH-NpLH2; and (4) Akaluc was brighter when paired with either CycLuc1 or Akalumine-HCl when paired with D-LH2.
Conclusion: We believe that the experimental results described here should provide valuable guidance to end users for choosing the correct luciferin/luciferase pairs for a variety of BLI applications.
In this study, we provide numerous updates and additions to the checklist of click-beetles (Elateridae) of Iran based on the most up-to-date family classification as well as on the recent taxonomic studies dealing with the species diversity of the click-beetles from Iran. The updated checklist of Iranian Elateridae includes 293 species classified in 62 genera and seven subfamilies, i.e.
Chemical and Behavioral Ecology, Gembloux Agro-Bio Tech, University of Liege, Avenue de la Faculté d'Agronomie 2B, Gembloux, 5030, Belgium. Electronic address:
Entomopathogenic nematodes (EPNs) have emerged as a promising tool for controlling soil-dwelling crop pests. However, their efficacy varies according to EPN populations and targeted hosts. Wireworms are polyphagous insects causing significant crop losses, especially since the ban on pesticides previously used for their control.
Wireworms, the larva of click beetle (Agriotes species), are one of the most destructive pests of horticultural crops in North America, responsible for considerable economic losses in Canada. Agriotes sputator (A. sputator) species is a predominant wireworm pest attacking potato fields in Eastern Canada.
The revocation of chlorpyrifos tolerances in 2022 left sweetpotato growers without their most important tool to combat a complex of soil-borne root pests that includes wireworms (Coleoptera: Elateridae). Since then, growers have reported increased root damage despite a rapid pivot to pyrethroid-based management systems to replace mechanically incorporated preplant chlorpyrifos broadcast sprays. Our goal was to evaluate the efficacy of alternative insecticide chemistries and application methods to expand the portfolio of management options for wireworms, specifically Conoderus spp.
