siRNA screening for gene silencing in motor neurons derived embryonic stem cells (ES) using DreamFect Gold transfection reagent

DreamFect Gold was chosen among eight transfection reagents to conduct siRNA screening apporach due to its high cell viability score and effective knockdown,. A library of 133 genes was screened for siRNA silencing in motor neurons derived ES using DreamFect Gold.

This article demonstrates the high efficiency of DreamFect Gold from OZ Biosciences for siRNA screening and gene silencing in motor neurons derived ES.

article reference: Front Cell Neurosci. 2014 May 20;8:140. doi: 10.3389/fncel.2014.00140. eCollection 2014.

siRNA screen of ES cell-derived motor neurons identifies novel regulators of tetanus toxin and neurotrophin receptor trafficking.

Terenzio M, Golding M, Schiavo G.

Abstract

Neurons rely on the long-range transport of several signaling molecules such as neurotrophins and their receptors, which are required for neuronal development, function and survival. However, the nature of the machinery controlling the trafficking of signaling endosomes containing activated neurotrophin receptors has not yet been completely elucidated. We aimed to identify new players involved in the dynamics of neurotrophin signaling endosomes using a medium-throughput unbiased siRNA screening approach to quantify the intracellular accumulation of two fluorescently tagged reporters: the binding fragment of tetanus neurotoxin (HCT), and an antibody directed against the neurotrophin receptor p75(NTR). This screen performed in motor neurons differentiated from mouse embryonic stem (ES) cells identified a number of candidate genes encoding molecular motors and motor adaptor proteins involved in regulating the intracellular trafficking of these probes. Bicaudal D homolog 1 (BICD1), a molecular motor adaptor with pleiotropic roles in intracellular trafficking, was selected for further analyses, which revealed that BICD1 regulates the intracellular trafficking of HCT and neurotrophin receptors and likely plays an important role in nervous system development and function.

DreamFect™Gold allows transfecting all types of nucleic acids with a very high efficiency. Due to its formulation, DreamFect™Gold delivers a large quantity of nucleic acids leading to higher protein expression compared to other transfection reagents. It is fully biodegradable and does not interfere with cellular mechanisms.