A Bright Future with a Small, Versatile Luciferase

Promega Connections is pleased to present this guest blog from Kyle Hooper, PhD, Cellular Analysis Specialist in the North America Group at Promega Corporation

NanoLuc™ Luciferase has properties that make it especially useful for imaging. Here a beta-2 adrenergic receptor/NLuc fusion is directed to the cell surface.

Check out the new webinar about NanoLuc™ Luciferase just posted to the Promega Webinars site. NanoLuc™ Luciferase is derived from a deep sea shrimp and is ~100-fold brighter than either firefly or Renilla luciferase. The first part of the webinar reviews the generation on NanoLuc™ Luciferase and its substrate, furimazine, as well as its use as an intracellular genetic reporter. NanoLuc™ Luciferase can also be secreted from the cell and allows kinetic studies on the same cells without cell lysis. Simply remove a portion of the conditioned cell culture medium and perform the luciferase assay. NanoLuc™ Luciferase is a useful tool in the role of a traditional genetic reporter and the brightness makes it more sensitive than any other reporter. The increased brightness could compensate for poor transfection efficiency allowing you to move assays into more biologically relevant cell lines from easy-to-transfect, less relevant cell lines. Perhaps the bright signal could move that assay from a 12-well plate to a 96-well plate or a 96-well assay to a 384-well assay.

BRET is one of the exciting new applications for the NanoLuc™ Luciferase.

Perhaps the most exciting application of NanoLuc™ Luciferase is its ability to function as a protein fusion tag. NanoLuc™ Luciferase is 19kDa, smaller than many fusion tags and one third the size of firefly luciferase. The seminar examines how tagged proteins can be followed within cells through bioluminescent imaging. Don’t miss the movie of NanoLuc™ Luciferase-tagged glucocorticoid receptor moving from the cytoplasm to the nucleus upon binding dexamethasone. We also see how NanoLuc™ Luciferase fusions can be used to monitor the stability of a protein through examination of a p53 model system. An application that gets a lot of interest is the potential use NanoLuc™ Luciferase potential as the luminescent donor in bioluminescent resonance energy transfer or BRET. We demonstrate how NanoLuc™ Luciferase and our other fusion tag, HaloTag® Fusion Protein, can be used to make an excellent BRET pair and application to a common BRET model system to investigate protein:protein interactions. I hope you agree, the future for NanoLuc™ Luciferase is indeed bright.

For Further Reading

  1. Hall, M.P. et al. (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem. Biol. epub ahead of print.
  2. NanoLuc™: A Smaller, Brighter, More Stable Luciferase Reporter (poster)
  3. NanoLuc™ Lucferase Technology Overview

About the Webinar Series
www.promega.com/webinars provides a schedule of upcoming webinars. In addition there are links to previous webinars that allow you to either view the recording or download a pdf of the presentation. There is also a pdf of additional material available for each past webinar.

To register for a webinar, use the “Registration” link at www.promega.com/webinars This allows you to not only view the webinar, but also to participate in the live chat.

Need a reminder? You can also sign-up for monthly invitations to webinars at the webinars page (see link above). Note: Live chat is only available for live webinars, not links to recorded webinars.

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Promega products are used by life scientists who are asking fundamental questions about biological processes and by scientists who are applying scientific knowledge to diagnose and treat diseases, discover new therapeutics, and use genetics and DNA testing for human identification. Originally, founded in 1978 in Madison, Wisconsin, USA, Promega has branches in 16 countries and more than global distributors serving 100 countries.

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