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Application of RNAi and heat-shock-induced transcription factor expression to reprogram germ cells to neurons in C. elegans

Item Type:Article
Title:Application of RNAi and heat-shock-induced transcription factor expression to reprogram germ cells to neurons in C. elegans
Creators Name:Kolundzic, E., Seelk, S. and Tursun, B.
Abstract:Studying the cell biological processes during converting the identities of specific cell types provides important insights into mechanism that maintain and protect cellular identities. The conversion of germ cells into specific neurons in the nematode Caenorhabditis elegans (C. elegans) is a powerful tool for performing genetic screens in order to dissect regulatory pathways that safeguard established cell identities. Reprogramming of germ cells to a specific type of neurons termed ASE requires transgenic animals that allow broad over-expression of the Zn-finger transcription factor (TF) CHE-1. Endogenous CHE-1 is expressed exclusively in two head neurons and is required to specify the glutamatergic ASE neurons fate, which can easily be visualized by the gcy-5prom::gfp reporter. A trans gene containing the heat-shock promoter-driven che-1 gene expression construct allows broad mis-expression of CHE-1 in the entire animal upon heat-shock treatment. The combination of RNAi against the chromatin-regulating factor LIN-53 and heat-shock-induced che-1 over-expression leads to reprogramming of germ cell into ASE neuron-like cells. We describe here the specific RNAi procedure and appropriate conditions for heat-shock treatment of transgenic animals in order to successfully induce germ cell to neuron conversion.
Keywords:Developmental Biology, Germ Cell, Transcription Factor-Induced Reprogramming, Epigenetic, Reprogramming Barrier, RNA Interference, Heat-Shock Induction, Neuronal Reprogramming, Animals, Caenorhabditis elegans
Source:Journal of Visualized Experiments
Page Range:e56889
Date:1 January 2018
Official Publication:https://doi.org/10.3791/56889
External Fulltext:View full text on PubMed Central
PubMed:View item in PubMed

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