Cell Fate Reprogramming of Nematode Germ Cells into Neurons
Transcrição
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.
