Cellecta - Pre-designed Validated shRNA Libraries

Pre-designed Validated shRNA Libraries

At the heart of Cellecta's technological repertoire is the pooled lentiviral shRNA library. Using our innovative method which allows us to quickly and cost-effectively construct a library, we are capable of generating libraries of any size. Combined with our proprietary vector and shRNA designs, our pre-designed validated pooled shRNA libraries are not only cost-effective, they are also the most powerful and effective HT RNAi screening tools available on the market.

The second generation validated pooled shRNA libraries, launched in April of 2010, were created using proprietary software for in silico prediction of functional shRNAs, a database of validated shRNA, and optimized designs of both expression vectors and shRNA. The vector and shRNA design alone delivers approximately 55-70% functional shRNA (>70% knockdown) depending on the cell type. In comparison, the percentages of functional shRNA for two other major shRNA library providers are 50% (21-mer shRNA design) and 20% (miR30 design). The experiment and data generated using our first Validated Druggable Genome 25K shRNA Library is described on the HT RNAi TGF-β Screen Comparison Data page.

A result of over 5 years of NIH-funded research (SBIR Grant numbers 5R44HG003355 and 1R43RR024095), our "pre-designed" pools of validated shRNAs are ready-for-ligation to any of our vectors to produce customized validated shRNA libraries. The table below summarizes Cellecta's new Pre-designed, Validated Pooled Lentiviral Bar-Coded shRNA Libraries.

Pre-Designed Validated shRNA Libraries (Human and Mouse)	# of target mRNAs	shRNA complexity
Druggable Genome	8,000	25,000
Signaling Pathways	5,000	27,000
Cancer-Specific	5,000	27,000
Disease-Specific	5,000	27,000

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.

diagram of shRNA Library construction and HT RNAi screen

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.

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.