Custom Pooled Lentiviral Bar-coded shRNA Libraries
Technology
Functional Data & Presentations
At the heart of Cellecta's technological repertoire is the pooled lentiviral bar-coded shRNA library. Using an innovative method which allows the rapid construction of a pooled library, libraries of any size can be generated on a custom basis. The quality of the library is not compromised by the method—our proprietary vector and shRNA design alone delivers approximately 55-70% functional shRNA (>70% knockdown) depending on the cell type.
In 1 to 4 months, we can provide you with the oligo, plasmid, or plasmid + packaged pooled shRNA library designed to target your set of genes. Choose any fluorescent marker and antibiotic resistance gene, and a customized dual marker lentivector will be developed for you if we don't already have it. We will provide the list of shRNA sequences, vector sequence, customized protocols, and as much technical support you need to successfully plan and execute a functional screen.
The advantages of using Cellecta's screening method over other methods include:
- Low-cost technology for construction of focused or genome-wide pooled shRNA lentiviral libraries.
- Second generation pre-designed validated shRNA libraries based on proprietary software for in silico prediction of functional shRNAs, a database of validated shRNA, and optimized designs of both expression vectors and shRNA.
- A highly optimized shRNA structure increases target knockdown efficiency and reduces percentage of false negative hits.
- HT sequencing detection of shRNA-specific bar-codes improves positive identification of shRNAs which modulate phenotypic responses.
- Tet-regulatable shRNA expression enhances efficiency of shRNA enrichment.
- Use of lentiviral vectors enables efficient delivery of high complexity, pooled shRNA libraries into a wide range of cell types.
- Cellecta shRNA libraries can also be designed for compatibility with custom microarrays, however we highly recommend HT sequencing over hybridization.
For ordering information, visit the Custom shRNA Library Order page.
Pooled Lentiviral Bar-Coded shRNA Library Overview
In the pooled format strategy, a lentiviral packaged library encoding a pooled set of bar-coded 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 HT sequencing of shRNA-specific bar-codes.
Fig. 1. Flowchart of Cellecta's HT RNAi Screen services, including construction of pooled bar-coded shRNA libraries, functional screen and phenotypic selection, and bioinformatic data and pathway analysis.
Oligo Pools Now Available
We can now deliver pre-designed shRNA libraries as purified template oligos in pooled format. Oligos are provided as single-strand templates—customer must perform PCR, restriction digest, and PAGE purification. Alternatively, Cellecta can provide this service for an additional fee. We can do as much additional customization as necessary. Customer vector must have cohesive ends compatible to the default configuration of the pre-designed shRNA oligos. Please contact us for additional information.
Valuable Resources Online
View our Frequently Asked Questions (FAQs) page for detailed information on our pooled lentiviral shRNA libraries. For relevant references and literature, visit the Articles page under Resources. View the TGF-β and Prostate Cancer Viability screen experimental data pages for examples of HT RNAi screens.
Please read the Cellecta-Agilent License Statement for the terms of use.
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