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Custom Functionally-Validated shRNA Libraries

Scientists at Cellecta have developed a novel high-throughput (HT), cell-based reporter assay for unbiased selection of functional shRNAs that effectively knock down target genes. Functional shRNAs can be identified for a customer-supplied list of target genes or shRNAs, and we can generate lentiviral libraries of the functionally validated (FV) shRNAs. A series of validation screens is carried out to determine the best knockdown candidates, while at the same time care is taken to minimize any off-target effects. For ordering information, visit our Custom shRNA Library Order page. For more information about our functional validation method, visit the shRNA Validation Technology page. Cellecta also offers Pre-Designed FV shRNA libraries.

The advantages of using Cellecta's screening method over other methods include:

  • Proprietary ultra-high throughput oligonucleotide synthesis (chip-based) for construction of genome-wide shRNA libraries.
  • Second generation of functionally validated siRNA libraries based on proprietary ultra-high throughput validation technology.
  • Use of Lentiviral vectors for efficient delivery of high complexity, pooled siRNA libraries into cells.
  • Cellecta libraries are bar-coded, and they can be used with custom arrays (customer can order from Agilent Technologies) or with HT sequencing (Cellecta can provide at additional cost) to identify shRNA which modulate phenotypic responses.

In the pooled format strategy, a library encoding a pooled set of packaged viral shRNA constructs is introduced into a population of identical cells under conditions where each transduced cell will express only a single gene-specific siRNA. Cells exhibiting the desired phenotypic changes are isolated and the shRNA constructs, presumably inducing the phenotypes, are recovered by PCR and identified by sequence analysis or microarray hybridization using bar-coding strategy, or designing siRNAs complementary to existing commercial microarrays.

An example of a typical experiment and resulting functional data is available in our High Throughput RNAi Screen data section. View our Frequently Asked Questions (FAQs) page for detailed information on our pooled lentiviral shRNA libraries.