JoVE Encyclopedia of Experiments
Neuroscience
0 views • 2:59 min • July 8th, 2025
Take genetically engineered bacteria on a nematode growth medium.
The bacteria carry a plasmid with a nematode chromatin-regulating factor gene.
The medium contains an inducer that triggers the expression of a transgenic RNA polymerase from the bacterial genome that transcribes the chromatin-regulating factor gene into a dsRNA.
Add transgenic nematode larvae carrying a neuron fate-inducing transcription factor gene under a heat-shock promoter.
Upon bacterial consumption, the ingested dsRNA generates siRNA that downregulates the chromatin-regulating factor expression, facilitating cell fate reprogramming.
Through the parents' germ cells, the siRNA is passed onto the progeny.
Downregulation of the chromatin-regulating factor expression in the progeny germ cells, indicated by a protruding vulva phenotype, allows their reprogramming.
Apply heat shock to the progeny, activating the heat-shock promoter and expressing the neuron fate-inducing transcription factor that triggers neuronal differentiation, transforming germ cells into neurons.
The neurons express a transgenic fluorescent reporter protein, facilitating microscopic visualization.
Manually transfer 50 L4 worms per replicate to a
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