OZ Biosciences Blog

Tuesday, December 2, 2014

Gene Silencing in MCF-10A with siRNA using Lullaby transfection reagent

MCF10A were transfected with siRNA oligo at a final concentration of 37.5 nM using Lullaby according to the protocol recommendations.

This article demonstrates the high efficiency of Lullaby, a transfection reagent from OZ Biosciences dedicated to siRNA to induce gene silencing into mammary gland epithelial cells.

article reference: Nat Cell Biol. 2014 Nov;16(11):1092-104.

ASPP2 controls epithelial plasticity and inhibits metastasis through β-catenin-dependent regulation of ZEB1.

Wang Y, Bu F, Royer C, Serres S, Larkin JR, Soto MS, Sibson NR, Salter V, Fritzsche F, Turnquist C, Koch S, Zak J, Zhong S, Wu G, Liang A, Olofsen PA, Moch H, Hancock DC, Downward J, Goldin RD, Zhao J, Tong X, Guo Y, Lu X.
Abstract
Epithelial to mesenchymal transition (EMT), and the reverse mesenchymal to epithelial transition (MET), are known examples of epithelial plasticity that are important in kidney development and cancer metastasis. Here we identify ASPP2, a haploinsufficient tumour suppressor, p53 activator and PAR3 binding partner, as a molecular switch of MET and EMT. ASPP2 contributes to MET in mouse kidney in vivo. Mechanistically, ASPP2 induces MET through its PAR3-binding amino-terminus, independently of p53 binding. ASPP2 prevents β-catenin from transactivating ZEB1, directly by forming an ASPP2-β-catenin-E-cadherin ternary complex and indirectly by inhibiting β-catenin's N-terminal phosphorylation to stabilize the β-catenin-E-cadherin complex. ASPP2 limits the pro-invasive property of oncogenic RAS and inhibits tumour metastasis in vivo. Reduced ASPP2 expression results in EMT, and is associated with poor survival in hepatocellular carcinoma and breast cancer patients. Hence, ASPP2 is a key regulator of epithelial plasticity that connects cell polarity to the suppression of WNT signalling, EMT and tumour metastasis.

Lullaby is the ideal siRNA transfection reagent for gene silencing reaching up to 90% gene silencing. It protects siRNA from extracellular degradation and maintains high viability due to its bio-degradable properties.


No comments:

Post a Comment