Western Blot – Multimedia
Poster
Intended transcriptional gene silencing with siRNA to the human Vascular Endothelial Growth Factor (VEGF) promoter results in VEGF repression through sequence-specific off-targeting
SiRNA designed to target the human VEGF gene promoter revealed a putative candidate for transcriptional gene silencing. However, mutation or deletion of the target site demonstrated that the observed VEGF knockdown was the result of a sequence-specific off-target effect. Bioinformatic and microarray analyses of genes implicated in VEGF transcriptional control followed by knockdown and Western blotting experiments presented one candidate, GRB2, as an unintended target of the VEGF promoter-specifi
Poster
LacZ Reporter Fusion Assay for Rapid and Easy Identification of Highly Efficient siRNA and its Delivery by Lentivirus into Suspension Cell Lines
The LacZ reporter fusion assay is an excellent method to precisely quantify knockdown efficiency of siRNA-sequences and can be corroborated by Western blot analysis. Efficient production of Lentiviruses and high transduction efficiency (up to 98%) were achieved on lymphoid B- and T-cells as demonstrated by FACS analysis and GFP expression. Successful knockdown effect with specific shRNA was observed in suspension cells three days after lentiviral infection.
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