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Assessing Gene Function with siRNA Libraries


Andrea Kroenke1, Anne Grabner1, Christoph Sachse1, Kathy Latham2, and Christophe Echeverri1
1Cenix BioScience GmbH, 2Ambion, Inc.

Sets of siRNAs focused on a specific gene class, also called siRNA libraries, have the capacity to greatly increase the pace of pathway analysis and functional genomics. Here is an example of the power of Ambion's Silencer Kinase siRNA Library, as demonstrated through preliminary screening experiments carried out by Cenix BioScience GmbH. This highly validated library includes nearly 1800 individual siRNAs targeting 597 human kinases. In this experiment, 178 siRNAs from the library, targeting 178 different kinases expressed in HeLa cells, were each tested to determine their effects on cellular proliferation and mitotic index. This screen identified several kinase genes that are involved in the cell cycle, confirmed the identify of some kinases known to be involved, and identified a few kinases that had not previously been reported to be involved.


Experimental Design
The Silencer Kinase siRNA Library includes more than 600 validated siRNAs, which have been shown to reduce target mRNA levels by >70% when transfected into HeLa cells at a concentration of 100 nM. Cenix BioScience chose 178 of these validated siRNAs, targeting 178 diff
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