Knockdown of p53 by Accell Self-delivering siRNA Causes Inhibition of p53-dependent DNA Damage Response in IMR-32 Neuroblastoma Cell Line and β-amyloid Toxicity in Rat Cortical Neurons
Poster Jun 12, 2015
Žaklina Strezoska, Tamara Seredenina1, Devin Leake, Annaleen Vermeulen
Neuroblastoma cell lines and primary neuronal cultures are commonly used as cellular model systems for studying cancer and neuronal development as well as being highly relevant models for the study of neurodegenerative diseases. However, most neuroblastoma cell lines and practically all primary neuronal cells suffer from low transfection efficiency due to the refractory nature of the cells to lipid-based transfection reagents. As such, application of siRNA for inducing RNA interference (RNAi), has limited utility in these cell types; thus limiting their usefulness for development of functional assays for screening and discovery of novel disease-relevant genes.
Dharmacon™ Accell™ siRNA enables efficient delivery in a wide range of cell lines and primary cells. Accell siRNA reagents carry a novel chemical modication pattern that facilitates the delivery of siRNA without a need for transfection reagents. To demonstrate the utility of Accell siRNA reagents in neuronal cells, the effects of the down-regulation of the tumor suppressor p53 was examined. This gene plays a pivotal role in mediating DNA damage-induced apoptosis as well as conferring a protective effect from β-amyloid peptide-induced neurotoxicity. Here we describe how application of Accell siRNA enabled the development of a high content screening assay in IMR-32 neuroblastoma cells and a whole culture cell viability assay in primary rat cortical neurons. The ability to modulate gene expression in neuronal cell lines and primary neurons using Accell siRNA opens new opportunities for functional genomic siRNA screens in the eld of neuroscience.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE
International Conference on Cell and Structural biology
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