Directing neuronal differentiation of primary neural progenitor cells by gene knockdown approach.

DNA and Cell Biology
Wei Ching LowSing Yian Chew

Abstract

Directing differentiation of neural stem/progenitor cells (NPCs) to produce functional neurons is a promising remedy for neural pathological conditions. The major challenge, however, lies in the effective and efficient generation of a sizable population of neurons. A potential strategy is to incorporate RNA interference (RNAi) during directed stem cell differentiation to recapitulate the complex cell-signaling cascades that often occurs during the process. In this study, in vitro silencing of RE1-silencing transcription factor (REST) was carried out using small-interfering RNAs (siRNAs) to evaluate the efficacy of combining REST knockdown with conventional differentiation approaches to enhance neurogenesis. While earlier studies have demonstrated enhanced neuronal lineage commitment from embryonic stem cells and mesenchymal stem cells upon REST knockdown, the effects of REST silencing during other stages of neural development have not been extensively evaluated. We hypothesize that REST knockdown would enhance NPC development to mature neurons and that induced REST silencing can serve as a potential biochemical approach to direct cell fate. Under nonspecific induction conditions, REST knockdown induced eightfold higher Tuj1 mRN...Continue Reading

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Citations

Jul 8, 2014·FEBS Letters·Christelle En Lin ChuaBor Luen Tang
Dec 1, 2015·International Journal of Pharmaceutics·E AndréC N Montero-Menei
Dec 3, 2014·Biomaterials·Winifred Wing Yiu YauSing Yian Chew

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Methods Mentioned

BETA
immunoprecipitation assay
transfection
gene knockdown
PCR
confocal microscopy

Software Mentioned

siREST
ImageJ

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