The objective of the RNAi Service is to provide convenient, cost effective services for siRNA/shRNA design, siRNA/shRNA delivery into target cells and knock-down validation to
City of Hope Cancer Center investigators.

The primary services of the RNAi Service:

• siRNA/shRNA design
• optimized siRNA/shRNA delivery into target cells using cationic lipids, electroporation or lentiviral vectors
• screening of siRNA/shRNA libraries
• quantitative PCR

siRNA Design
The RNAi laboratory will design 27mer siRNA duplexes based on the gene accession number provided to the core and send the siRNA sequences to the core user for approval. A minimum order of 2 siRNA duplexes is required. 

siRNA Delivery
One of two methods is employed to transfect siRNA duplexes into target cells:

1. cationic lipids that are optimized for siRNA transfection or
2. electroporation using an Amaxa Nucleofector Device for efficient transfection of primary and difficult-to-transfect cell lines

Cy3-labeled siRNAs are used to initially monitor (by FACS) and optimize the transfection efficiency. 

Quantative PCR
qPCR is the most convenient way of detecting siRNA-induced gene silencing. The RNAi laboratory will analyze the gene silencing effect after 24 or 48h. Gene-specific probes (Roche Universal Probe Library) are used for all qPCR assays