Catalog #: CPLVSHL
Pooled lentiviral-based libraries containing heterogeneous mixtures of shRNA allow you to assay the effects of many thousands of pooled constructs in one experiment. By taking advantage of array-based oligonucleotide synthesis, we can readily make precisely defined large custom pooled libraries expressing many thousands of shRNAs. In approximately 2 months, we can produce a completely new, high-quality custom library focusing on any target set of genes. The library vector can also be easily customized, and the same library oligonucleotides can be cloned into more than one vector backbone (e.g. with an inducible and constitutive shRNA). It is a very flexible platform.
Screening readout of enrichment or depletion representation of particular shRNAs is greatly facilitated by the incorporation of easily sequenced barcodes in each shRNA construct. Direct amplification and sequencing of hairpins is problematic, and increase variability in the data since small sequence variations of the hairpin stem significantly affect the efficiency of PCR.
We can design libraries to target each transcript with as many shRNAs as desired. Given that shRNA can often have off-target effects due to its similarity with miRNA regulator sequences, we usually design shRNA libraries with 10 or more hairpins to each specific transcript. More than two-thirds of our optimized shRNAs knock down transcript levels by greater than 70%.