Primary hippocampal neuron transfection and primary cortical neuron transfection with oligonucleotides using NeuroMag

Rat primary hippocampal and cortical neurons were transfected at 14 to 21 DIV with 50nm Oligonucleotides using NeuroMag transfection reagent. 

This paper shows the high efficiency of NeuroMag transfection reagent from OZ Biosciences to transfect primary hippocampal and cortical neurons with miRNA.

paper reference: Hum Mol Genet. 2013 Apr 11.

De-repression of FOXO3a death axis by microRNA-132 and -212 causes neuronal apoptosis in Alzheimer's disease.
Wong HK, Veremeyko T, Patel N, Lemere CA, Walsh DM, Esau C, Vanderburg C, Krichevsky AM.

Abstract

Alzheimer's disease (AD) is a multifactorial and fatal neurodegenerative disorder for which the mechanisms leading to profound neuronal loss are incompletely recognized. MicroRNAs (miRNAs) are recently discovered small regulatory RNA molecules that repress gene expression and are increasingly acknowledged as prime regulators involved in human brain pathologies. Here we identified two homologous miRNAs, miR-132 and miR-212, down-regulated in temporal cortical areas and CA1 hippocampal neurons of human AD brains. Sequence-specific inhibition of miR-132 and miR-212 induces apoptosis in cultured primary neurons, whereas their overexpression is neuroprotective against oxidative stress. Using primary neurons and PC12 cells, we demonstrate that miR-132/212 controls cell survival by direct regulation of PTEN, FOXO3a, and P300, which are all key elements of AKT signaling pathway. Silencing of these three target genes by RNAi abrogates apoptosis caused by the miR-132/212 inhibition. We further demonstrate that mRNA and protein levels of PTEN, FOXO3a, P300, and most of the direct pro-apoptotic transcriptional targets of FOXO3a are significantly elevated in human AD brains. These results indicate that the miR-132/miR-212/PTEN/FOXO3a signaling pathway contributes to AD neurodegeneration.

NeuroMag is the first dedicated Magnetofection ™ transfection reagent for neurons. It is perfect for primary neurons and can also be used with cell lines and glial cells. The transfection can be performed with neurons from 3 DIV to 21 DIV. It can also be used on ES cell derived motor neurons.