OmicsLink shRNA clone collections include lentiviral and non-viral vector-based shRNA constructs against genome-wide human, mouse and rat genes. shRNA of varying lengths (19 to 29 bases) were designed using a proprietary algorithm to make shRNA expression constructs that have high knockdown efficiency with minimal off-target effect.
A set of four expression constructs is offered against every target gene with the guarantee that at least one of the four will have a knockdown effect of 70% or more on corresponding gene expression as determined by qRT-PCR. Refer to warranty and cancellation policy here.
All cell types covered
Lentiviral and non-viral vector option allows transfection or transduction of shRNA into difficult-to-transfect cells as well conventional dividing cell lines.
Multiple delivery formats
Markers and reporters
Vectors with mCherry or eGFP reporter gene for monitoring transfection or transduction efficiencies. Stable cell selection with puromycin marker.
Fully sequenced expression cassettes
Expression cassettes of all shRNA clones are fully sequenced including the promoter, sense and antisense target sequences, hairpin, termination and other linker sequences.
Figure 1. Lentiviral and non-viral expression vector-based shRNA clones
OmicsLink shRNA vectors
|Vector ||Promoter ||Selection Marker ||Reporter Gene ||Viral type |
|psiLv-H1 ||H1 ||Puromycin ||eGFP ||Lenti |
|psiLv-U6 ||U6 ||Puromycin ||eGFP ||Lenti |
|psiLv-mH1 ||H1 ||Puromycin ||mCherry ||Lenti |
|psiLv-mU6 ||U6 ||Puromycin ||mCherry ||Lenti |
|psi-H1 ||H1 ||Puromycin ||eGFP ||N/A |
|psi-U6 ||U6 ||Puromycin ||eGFP ||N/A |
|psi-mH1 ||H1 ||Puromycin ||mCherry ||N/A |
|psi-mU6 ||U6 ||Puromycin ||mCherry ||N/A |
|psi-nH1 ||H1 ||Puromycin ||N/A ||N/A |
|psi-nU6 ||U6 ||Puromycin ||N/A ||N/A |
|psiLv-nH1 ||H1 ||Puromycin ||N/A ||Lenti |
|psiLv-nU6 ||U6 ||Puromycin ||N/A ||Lenti |
|psiHIV-nH1 ||H1 ||Puromycin ||N/A ||Lenti |
|psiHIV-nU6 ||U6 ||Puromycin ||N/A ||Lenti |
Single gene down-regulation — The knocking down (KD) effect of the shRNA clones for a single gene can be studied and compared with that of a scrambled nucleotide control clone which is included for free with every shRNA clone order.
Pathway analysis — Genes have been grouped into various signal transduction, metabolic, and disease pathways and associations, as well as gene families and groups. By arraying the shRNA clones of known pathway(s) in 96 or 384 well plates, the role for a group of genes can be studied in a pathway. Large collection of shRNA clones against the human kinome is available which can be grouped according to the functionality of each kinase.
Figure 2. Mechanism of shRNA vector-mediated gene silencing
Measurement of knockdown (KD) effect by vector-based shRNA clones
Western blot analysis — Protein down-regulation is required in most cases for gene KD studies which can be assessed by western blot analysis.
Functional assays — KD effect of shRNA can be studied by end-point biological and biochemical assays such as cell proliferation, colony formation, cell cycle analysis, and migration based on the potential role played by each gene /protein.
Functional assays by a reporter gene/protein — Alternatively, when the endogenous transcript level of gene of interest is low and/or when qRT-PCR and western blot analysis is not feasible or possible, KD effect can be measured by co-transduction of shRNA clone with an expression clone plasmid which is transcribed into a chimeric mRNA transcript consisting of a reporter gene and the target gene ORF. The reporter gene encodes a reporter protein enzyme that can be used in functional assays. The destruction of the target gene mRNA by targeting shRNA results in the degradation of the reporter gene and, therefore, translation reduction of reporter protein enzyme. The KD effect can be quantified by the reporter enzymatic assays as illustrated by an example in figure 3 below.
Figure 3. Validation of shRNA against human kinome members using vector-mediated (psi-sH1) shRNAs and CMV driven expression clones (in psiCheck-AP) that contain target kinase ORFs and express alkaline phosphatase (AP) for colorimetric assays. A. In the absence of shRNA, AP is expressed and its activity is quantified by colorimetric assay. B. Co-transfection of shRNA and AP-ORF expression clones, chimeric mRNA for AP-ORF is destroyed and, subsequently AP translation and its activity is reduced.
Related products and services
Validated human kinome shRNA clone collections