shRNA Libraries & Constructs
Lentiviral shRNA Libraries & Constructs
Efficient and Specific Gene Knockdown
Pooled Lentiviral Human and Mouse shRNA Libraries
Pre-made pooled and custom lentiviral-based libraries containing heterogeneous mixtures of shRNA that allow you to assay the effects of many thousands of constructs in one experiment are provided. The pre-made, pooled lentiviral genome-wide shRNA libraries target nearly all protein-encoding genes for human or mouse. They have been used in a number of highly cited and published genetic screening experiments for discovery and functional characterization of novel therapeutic targets. In addition, precisely defined high quality custom pooled libraries with well designed shRNAs targeting any set of your genes of interest can be made.
- Easily obtain proven high quality, highly cited pooled shRNA libraries
- Confidently screen for genes functionally required for drug mechanisms of action, disease progression or other biological responses
- Embedded shRNA barcodes eliminate the need to sequence hairpins
Barcodes
Screening readout of enrichment or depletion representation of particular shRNAs is greatly facilitated by the incorporation of easily sequenced unambiguous barcodes in each shRNA construct. Direct amplification and sequencing of hairpins is problematic, and increases variability in the data since small sequence variations of the hairpin stem significantly affect the efficiency of PCR leading to increased noise in the data readout. The incorporation of shRNA-specific barcodes addresses this problem inherent to pooled shRNA screens.
Genetic Screen using a Pooled Lentiviral shRNA Library
Outline of primary shRNA library screen with pooled Cellecta lentiviral libraries.
shRNA Cloning and Expression Vectors & Control Constructs
Short double-stranded RNAs with sizes 19-29 bp can efficiently mediate gene silencing in mammalian cells by guiding sequence-specific degradation of target mRNA sequences. Synthetic double-stranded siRNA molecules can be introduced into cells to suppress gene expression transiently. Alternatively, shRNA templates can be cloned into an shRNA expression vector and expressed in the cells of choice.
Lentiviral expression vectors are the most effective vehicles for delivering genetic material to almost any mammalian cell - including non-dividing cells and whole model organisms. As with standard plasmid vectors, it is possible to introduce shRNA lentivector constructs in plasmid form into the cells with low-to-medium efficiency using conventional transfection protocols. However, by packaging the lentiviral shRNA construct in pseudoviral particles, you can obtain highly efficient transduction and heritable expression of siRNA - even with most difficult to transfect cells, like primary, stem, and differentiated cells. The expression construct transduced in cells is integrated into genomic DNA and provides stable, long-term expression of the target gene. Endogenously expressed siRNA effectors provide long-term silencing of the target gene and allow the researcher to generate cell lines and transgenic organisms with a stable knockdown phenotype for functional studies.
A large number of shRNA cloning vectors with constitutive or inducible versions of H1 or U6 shRNA promoters and different marker options like GFP, RFP, PuroR, BleoR, NeoR, or Hygro-HK are offered.
Custom shRNA cloning and shRNA library cloning services are also provided.