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SureSilencing shRNA Plasmids

For knockdown of human, mouse, or rat gene expression by RNA interference

  • 70% gene knockdown by at least 2 different shRNAs
  • Control shRNA plasmids for nonspecific and off-target effects
  • Multiple selection markers
  • Renewable shRNA resource
  • Verified control shRNAs available

SureSilencing shRNA Plasmids are available for every gene in the human, mouse, and rat genome. For each gene, a set of 4 shRNA plasmids are provided, each targeting a different section of the gene, to knock down the expression of the target gene using short-hairpin RNA.

Search in GeneGlobe
You can search for the following terms:
  • Entrez Gene IDs (e.g., 835)
  • RefSeq IDs (e.g., NM_032983, NP_116765)
  • Gene symbols (e.g., CASP2)
  • Cat. no. (e.g., SI00299551, QT01342509)
  • Sanger ID or Accession (e.g., hsa-let-7b, MI0000063)
  • CpG loci identification numbers (CG#) (e.g., CG17753661)

Do not enter species information in the Search box. Use the drop-down list to select your species of interest.
When searching for miRNAs, do not omit the hyphens. Use hyphens or spaces (e.g., search for hsa-let-7b or hsa let 7b. Do not search for hsalet7b).

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Product Product no. Cat. no. List price:
SureSilencing shRNA Plasmids, GFP
SureSilencing shPlasmid, GFP
336311 Varies
$957.00
Enter your targets in search box above.
SureSilencing shRNA Plasmids, Hygromycin
SureSilencing shPlasmid, Hygromycin
336312 Varies
$957.00
Enter your targets in search box above.
SureSilencing shRNA Plasmids, Neomycin
SureSilencing shPlasmid, Neomycin
336313 Varies
$957.00
Enter your targets in search box above.
SureSilencing shRNA Plasmids, Puromycin
SureSilencing shPlasmid, Puromycin
336314 Varies
$957.00
Enter your targets in search box above.

SureSilencing shRNA Plasmids are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


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Knockdown of gene expression by at least 70%.
SureSilencing shRNA Plasmids with GFP marker were transfected into HEK-293 cells. The transfection efficiency was estimated by fluorescence microscopy. The relative expression level of each target gene, normalized to transfection efficiency, was determined by quantitative, real-time RT-PCR using RT2 qPCR Primer Assays and plotted with the standard deviation of the biological and technical replicates.
Performance

SureSilencing shRNA Plasmids specifically knock down the expression of every human, mouse, or rat gene by RNA interference. For each gene, 4 separate shRNA designs are provided, packaged in a plasmid backbone.

The experimentally verified shRNA design algorithm ensures maximum gene specificity and RNAi efficacy. Greater than 70% knockdown for the targeted gene will be observed with at least 2 of the 4 shRNA plasmids (as measured by real-time RT-PCR in transfected cells upon FACS-based enrichment for GFP expression or selection for antibiotic resistance) (see figure "Knockdown of gene expression by at least 70%"). The availability of 2 effective sequences controls for nonspecific and off-target effects.

Principle

Short-hairpin RNA (shRNA), one method of enabling RNA interference (RNAi), uses a plasmid- or viral- based expression vector, delivered to a cell population by transfection. The short RNA sequence expressed by the vector folds into a short hairpin and is processed by the cellular RNAi machinery. SureSilencing shRNA Plasmids use a plasmid-based expression vector to enable RNAi.

SureSilencing shRNA Plasmids are available in 4 different plasmid formats. Plasmids contain either the hygromycin, neomycin, or puromycin resistance gene or GFP.

Plasmids containing the hygromycin, neomycin, or puromycin resistance gene enable selection of stably transfected cells and are well suited for the study of the long-term effects of gene knockdown in cells. Plasmids containing the GFP gene offer an alternative to siRNA, enabling the identification and tracking of transiently transfected cells. They are highly suited to the study of short-term gene knockdown effects. The ability to track or stably express shRNA enables RNAi in difficult-to-transfect cells.

Unlike chemically synthesized siRNA, plasmid-based shRNA provides a renewable source of RNAi. Enough plasmid can be amplified in the lab to complete a project. SureSilencing shRNA Plasmids also include complete shRNA sequence information.

Procedure

SureSilencing shRNA Plasmids are first transformed into competent E. coli cells. Colonies are picked for large-scale bacterial culture and large-scale, high-quality transfection-grade plasmid preparation. Gene-specific and negative control shRNA plasmids are transfected  into replicate wells of the cell line of interest and incubated for 24–48 hours.

Following transfection with a GFP SureSilencing shRNA Plasmid, cells can be tracked and characterized using fluorescence microscopy or enriched using fluorescence-activated cell sorting (FACS) or antibiotic selection. Following transfection with a hygromycin, neomycin, or puromycin SureSilencing shRNA Plasmid, cells may then be replated into medium containing the appropriate antibiotic to begin the process of selecting for and cloning stably transfected cells. The extent of knockdown can be verified using RT2 Profiler PCR Arrays or RT2 qPCR Primer Assays.

Applications

SureSilencing shRNA Plasmids are highly suited for gene function studies, removal of protein activity, and signaling pathway studies.

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Kit Handbooks (1)
For genomewide, plasmid-based RNA interference
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Instrument Technical Documents (1)
For efficient knockdown of every human, mouse, and rat gene
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