Excellent knockdown of intracellular trafficking proteins in human cell lines

University of Michigan
Pharmaceutical Sciences
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Life Technologies

Product Name:

RNAiMax

Catalog Number:

13778075

Our research explores the interaction of drug and gene delivery vehicles with intracellular trafficking processes such as those associated with endosomal and cholesterol trafficking. In particular, we were interested in transiently knocking down these trafficking proteins such as clathrin, caveolin, ARF6, flotillin amongst others. We used RNAimax as the liposomal delivery agent for the corresponding siRNA for these proteins and got excellent knockdown of these proteins

Experimental Design and Results Summary

Application

Gene Delivery Studies

Starting Material

HeLa cells, corresponding custom-ordered siRNA from Dharmacon

Protocol Overview

Simple incubation protocols were followed to form complexes between the siRNA and RNAiMax as detailed in the associated manual.

Tips

Dropwise addition of solutions helps

Results Summary

A representative image from our experiments is shown of the cell samples wherein we measured clathrin expression. The Western blot using the clathrin, caveolin, laminim (non-specific siRNA) and noRNA control siRNA clearly shows that siCLT(anti-clathrin siRNA) knocks down clathrin but the other siRNA have no effect on the clathrin expression.

Additional Notes

None

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Summary

The Good

Very specific knockdown. Easy to use and stores well.

The Bad

None

The Bottom Line

Easy knockdown of your target mRNA via the use RNAimax

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