Shining Stars: Cool NanoLuc® Plasmid Constructs Available Through the Addgene Repository

Researchers having been sharing plasmids ever since there were plasmids to share. Back when I was in the lab, if you read a paper and saw an interesting construct you wished to use, you could either make it yourself or you could “clone by phone”.  One of my professors was excellent at phone cloning with labs around the world and had specific strategies and tactics for getting the plasmids he wanted. Addgene makes this so much easier to share your constructs from lab to lab. Promega supports the Addgene mission statement: Accelerate research and discovery by improving access to useful research materials and information.  Many of our technology platforms like HaloTag® Fusion Protein, codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc® Luciferase are present in the repository. We encourage people to go to Addgene to get new innovative tools. Afterall, isn’t science better when we share?

I’d like to focus on some tools in the Addgene collection based on NanoLuc® Luciferase (NLuc).  The creation of NanoLuc® Luciferase and the optimal substrate furimazine is a good story (1).  From a deep sea shrimp to a compact powerhouse of bioluminescence, NLuc is 100-fold brighter than our more common luciferases like firefly (FLuc) and Renilla (RLuc) luciferase.  This is important not so much for how bright you can make a reaction but for how sensitive you can make a reaction.  NLuc requires 100-fold less protein to produce the same amount of light from a Fluc or RLuc reaction.  NLuc lets you work at physiological concentrations.  NLuc is bright enough to detect endogenous tagged genes generated through the CRISPR/Cas9 knock-in.  NLuc is very inviting for endogenous tagging as it is only 19kDa.  An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined in Schmid-Burgk, J.L. et al (2).

Two applications of NanoLuc® Technology have caught my attention through coupling the luciferase with fluorescent proteins to make better imaging reporters and biosensors. Continue reading

Prepublication: Everybody’s Doing It?

Imagine for a moment this conversation between a senior graduate student and his dissertation adviser:

“Everybody’s doing it. Physicists and computer scientists do it all the time. And even Carol Greider has done it, and she’s a Nobel laureate.”

“Yes,” his adviser from her work, “she is a Nobel laureate; she can take that risk. But, I don’t have tenure, and I am still working on my first NIH grant. You don’t have a degree yet. None of these things—your PhD, the grant renewal, my promotion—come without publications in a peer-reviewed journal, and most peer-reviewed journals in our field, at the least the ones that count for grant renewals and promotion, don’t allow publication of previously released data.”

“But why let the publishers decide what is good science—why not let the scientific community decide and crowd source the review?”

“I agree, but I also want a future. We write the paper and submit it. So do your homework, let’s go to a journal with a short turnaround time, open review, and a reputation for publishing good science.”

Open Data and the Biological Sciences

The debate over prepublication in biology is raging.  Prepublication is the standard in physics, computer science, math, and economics to get results publicly available quickly for scientific commentary, and it doesn’t seem to interfere with career advancement and grant renewals. Is there a good reason that the same practice isn’t followed in the life/biological sciences? Continue reading