Nucleofection of miRNA Substrates
MicroRNAs (miRNAs), endogenous, small, non-coding RNA molecules, are key regulators of gene expression at the level of translation. They are differentially expressed in tissues, critical in the development of organisms, involved in viral infection, and associated with oncogenesis.
As with any other gene classes, their functionality is analyzed by overexpression using transfection of synthetic miRNAs or miRNA-expressing plasmids or by down-regulation using transfection of miRNA inhibitors.
The Nucleofector Technology allows studying this gene class in difficult-to-transfect cell types including suspension cell lines and primary cells.
- More than 90% transfection efficiency for both, RNA oligonucleotides and DNA vectors even in suspension cell lines and primary cells
- 96-well Shuttle enabling miRNA library screening
Expression of miR132 induces neurite sprouting by targeting a protein that represses neurite outgrowth (p250GAP).
Rat neonatal cortical neurons were transfected with a GFP reporter (green) and co-transfected with vector control, or expression constructs for premiR1-1 or premiR132 using Rat Neuron Nucleofector Kit and Nucleofector Device. Cells were immunostained for the neuronal marker MAP2 (red). Only cells transfected with premiR132 show neurite sprouting.
Vo et al., reproduced from the Proc. Natl. Acad. Sci. USA, 102(45): 16429 by copyright of the National Academy of Science and by permission of the authors.
