Custom lentiviral pooled screening libraries offer a personalized and efficient method for screening large numbers of lentiviral sgRNAs, shRNAs, or microRNAs without the requirement of automation. When a Dharmacon catalog screening library does not meet your needs, we can help you design your own custom lentiviral pools. All you need to do it selecting the product format desired, the number of genes, constructs per gene, and species.
Optimized construction of high-quality pools, complete analysis tools, and validated protocols are essential to a successful outcome when screening with pooled lentiviral libraries. Whether you need to perform a CRISPR knockout screen with lentiviral sgRNAs, a knockdown RNAi screen with shRNA, or a gene regulation screen with expressed microRNAs, a custom library can be built to your experimental requirements.
Library construction, pooling techniques, and high-throughput sequencing compatible screening workflows have all been experimentally validated to ensure reproducibility and accurate hit identification. Pools are provided as concentrated lentiviral particles for transduction into dividing and non-dividing cells.
Experimentally validated protocols and calculation sheets are readily available to help you with every aspect of the experiment from determining relative titer in your specific cells to calculating exactly how much volume you will require to complete the experiment. Our Bioinformatic analysis protocol provides full instructions on using open source software to run the analysis from the NGS data.
To learn more about the critical parameters of successful pooled lentiviral screening, including the conditions necessary for maintaining a high fold-representation, please download the following publication: Ž. Strezoska, A. Licon, Optimized PCR Conditions and Increased shRNA Fold Representation Improve Reproducibility of Pooled shRNA Screens. PLoS One 7, e42341 (2012).
Titer: Constitutive vectors, 5 x 108 TU/mL (± 20%); inducible vectors, 1 x 107 TU/mL (± 20%); all products are purified lentiviral particles with functional titers determined by qPCR.
Pool size: minimum of 50 total constructs up to 12,000 constructs per pool
Volume: Minimum of 100 uL up to 5 mL per pool; delivered in 25 µL aliquots
Custom lentiviral pooled libraries can be generated from any of the following product lines:
Lentiviral sgRNA for human, mouse and rat designed with our proprietary algorithm for improved functional knockout and best-in-class specificity checking.
Advanced rationally-designed microRNA-based shRNA for human, mouse, and rat genes using our patented scaffold and design characteristics. Choose from multiple promoter and reporter options for ultimate experimental flexibility.
Innovative microRNA vector design coupled with your choice of optimal promoter and reporter for over-expression of mature microRNAs.
Efficient gene silencing with a microRNA-adapted shRNA design. Available as lentiviral vector constructs or high-titer lentiviral particles for human and mouse