LentiExpress: Faster And Easier High-Throughput RNAi Screening

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RNAi Screening

Sigma-Aldrich introduces the LentiExpress shRNA-based system for rapid high-throughput RNAi screening with minimal reagent preparation or optimization required.

The addition of this technology to the MISSION shRNA platform pairs the benefits of lentiviral-based shRNA with a simple streamlined protocol.

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The bottleneck for researchers is the time and expense required to generate viral particles and devise a robust screening strategy. With the MISSION LentiExpress technology, they can eliminate reagent preparation with a sophisticated yet simple method to perform complex screens. An Optimization Plate enables researchers to quickly optimize the system for their particular cell line. This optimization process is uniquely simple compared to cumbersome transfection optimization methods that exist for siRNA-based screens. A researcher simply adds the desired number of cells to each well, continues with optional selection and/or addition of small molecules (for example, pharmaceutical compounds), and then proceeds directly to the desired assay for gene silencing or loss-of-function.

The LentiExpress format with the Human Kinase shRNA collection of The RNAi Consortium (TRC) enables rapid human kinome RNAi screening. The MISSION LentiExpress Human Kinase shRNA set consists of 3,109 individual pre-arrayed lentiviral clones harboring shRNA sequences that target 673 human kinase genes for gene silencing.

"Because we created the product to be ready-to-go single-use assays, researchers now have access to a fast and affordable means to utilize RNAi in whole-kinome screens. Lentiviral-based shRNA is also known for the benefit of being able to deliver RNAi effector molecules to a wide variety of cells including primary and non-transfectable cells lines," said Dr. Edward Weinstein, Manager of Functional Genomics Operations at Sigma-Aldrich. "Another benefit is stable long-term knockdown compared to transient transfection based siRNA options."

"Sigma-Aldrich researchers have performed proof-of-principle LentiExpress screens examining which genes might play a role in the modulation of a well-known cancer chemotherapy drug," said Dr. David Smoller, President of the Sigma-Aldrich Research Biotech business unit. "The screen resulted in hits already validated by the literature as well as additional hits that could be further investigated as prognostic markers for susceptibility to treatment. Both internal and external validations of the methodology have proven the utility of this exciting technology."

The LentiExpress technology complements the already robust MISSION shRNA product line which includes libraries, gene family sets, and gene target sets for The RNAi Consortium lentiviral-based shRNA collection targeting the human and mouse genomes. The product portfolio includes a viral packaging system, a variety of controls, as well as the unique ability to offer any or all of the collection in a choice of glycerol stock, purified DNA or viral particle format. Sigma-Aldrich's complete RNAi platform also includes the MISSION siRNA libraries, mRNA detection reagents, antibodies and AQUA peptides for protein level detection as well as an entire workflow of supporting reagents (cell culture, cell biology compounds and assays, transfection reagents, and much more).

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