A recent visit by videographers from the Journal of Visualized Experiments (JoVE) to the laboratory of geneticist and neurobiologist Dan Chase in the Biochemistry and Molecular Biology Department highlighted special experimental techniques his lab uses to perform genome-wide RNA interference (RNAi) screens. “These RNAi screens can be used to identify novel genes involved in many important biological processes,” says Chase.
The Chase lab uses the RNAi knockdown approach to search for genes that mediate neuron excitability, particularly those genes that act in dopamine signaling pathways in the roundworm C. elegans. He says, “C. elegans is a very useful model organism for studying neurotransmission because it has only 302 neurons compared to the billions of neurons found in the mammalian brain. Despite its simplicity, however, the fundamental mechanisms of neurotransmission found in mammals are also found in the worm.”
“RNA interference screens are a powerful means to survey the genome for genes involved in nearly any biological process,” he adds. “However, most labs have trouble getting the knockdown strategy to work consistently, and as a result, the strategy is underutilized.”
“We also had trouble with this approach initially, so we deconstructed the screening process and discovered the cause of the reduced knockdown efficiency. Using our improved protocols we are now able to knockdown the function of even the most difficult genes. The editors of JoVE recognized this from our 2012 publication where we used RNAi screens to identify several new genes involved in neurotransmitter signaling. They invited us to publish our new protocol in their journal.”
JoVE came to campus May 22 with camera crews to visit the Chase laboratory. Based on the manuscript authored in the Chase lab, a group of JoVE scriptwriters, animators and videographers filmed the RNAi knockdown procedure and will release the video online this summer.
Postdoc Katie Maher and graduate student Mary Catanese from the Chase lab were on camera during the journal’s May 22 campus visit “because they worked hard to develop this improved protocol and they do it perfectly every time,” he points out.
The neurobiologist notes, “A lot of experimental techniques, most of them cutting-edge, are very difficult to perform well, and many are not described adequately in the literature. JoVE videotapes difficult-to-perform protocols. Often there are nuances to experimental techniques that you can only learn by seeing them performed by an expert. For RNAi interference screens, Katie and Mary are the experts. They figured out why this technique was failing and they fixed it. Now withJoVE publishing their improved RNAi knockdown technique, other research labs can benefit from their hard work.”
JoVEis the first peer-reviewed, PubMed-indexed scientific video journal with a goal to increase the productivity of scientific research. Publishers say the online resource presents “novel techniques, novel applications of existing techniques, and gold standard protocols” that will be useful to other scientists.
Photo: Postdoc Katie Maher and videographer Kae Jae