miRNA and LNA transfection into Be(2)C cell line using Magnetofection - NeuroMag

NeuroMag transfection reagent was used to transfect miRNA inhibitor and LNA into Be(2)C cells for 48H.

This paper shows the high efficiency of NeuroMag from OZ Biosciences to transfect neuroblastoma cells.

article reference: Mol Pharmacol. 2013 Dec 3. 

Morphine Regulates Expression of MOR-1A, an Intron-retention Carboxyl Terminal Splice Variant of the μ Opioid Receptor (OPRM1) Gene Via miR-103/miR-107.

Lu Z, Xu J, Xu M, Pasternak GW, Pan YX.

Abstract

The μ opioid receptor (MOR-1) gene, OPRM1, undergoes extensive alternative splicing, generating an array of splice variants. Of these variants, MOR-1A, an intron-retention carboxyl terminal splice variant identical to MOR-1 except for the terminal intracellular tail encoded by exon 3b, is quite abundant and conserved from rodent to humans. Increasing evidence indicates that miroRNAs (miRNAs) regulate MOR-1 expression and μ agonists such as morphine modulate miRNA expression. However, little is known about miRNA regulation of the OPRM1 splice variants. Using 3' rapid amplification cDNA end (RACE) and Northern blot analyses, the present study identified the complete 3' untranslated region (3'-UTR) for both mouse and human MOR-1A and their conserved polyadenylation site, and defined the role the 3'-UTR in mRNA stability using a luciferase reporter assay. Computer models predicted a conserved miR 103/107 targeting site in the 3'-UTR of both mouse and human MOR-1A. The functional relevance of miR-103/107 in regulating expression of MOR-1A protein through the consensus miR-103/107 binding sites in the 3'- UTR was established by using mutagenesis and a miR-107 inhibitor in transfected HEK293 cells and Be(2)C cells that endogenously express human MOR-1A. Chronic morphine treatment significantly upregulated miR-103 and miR-107 levels, leading to downregulation of polyribosome-associated MOR-1A in both Be(2)C cells and the striatum of a morphine tolerant mouse, providing a new perspective on understanding the roles of miRNAs and OPRM1 splice variants in modulating the complex actions of morphine in animals and humans.

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.