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miScript miRNA Mimics

For studies on miRNA function and gene regulation using synthetic miRNA

  • Ready-to-transfect mimics behave like endogenous miRNAs
  • Available for all human, mouse, and rat miRNAs in miRBase
  • Highly flexible formats, scales, and grades
  • Custom design available for miRNAs not listed in miRBase
  • Easy to order via GeneGlobe

miScript miRNA Mimics are chemically synthesized, double-stranded RNAs which mimic mature endogenous miRNAs after transfection into cells. miScript miRNA Mimics are available at cell-culture grade (>90% purity) or animal grade (HPLC purified; for in vivo applications). For flexible screening, miScript miRNA Mimic Plates enable researchers to order any predesigned or custom-designed miScript miRNA Mimics in their choice of layout in 96-well or 384-well plates. miScript miRNA Inhibitors are suitable for similar experiments. For transfection, QIAGEN provides HiPerFect Transfection Reagent for miRNA Mimic transfection and Attractene Transfection Reagent for miRNA Mimic-DNA cotransfection.

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:
miScript miRNA Mimic
1 nmol (cell-culture grade), 5 nmol (cell-culture grade), or 20 nmol (animal grade) miScript miRNA Mimic, provided in tubes
219600 Varies Enter your targets in search box above.

miScript miRNA Mimics are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


0
Comparable downregulation by endogenous miRNA and miScript miRNA Mimic.
miR-1 is not endogenously expressed in HeLa S3 cells. miR-16 is endogenously expressed at high levels in HeLa S3 cells (Lagos-Quintana, M. et al. [2001] Identification of novel genes coding for small expressed RNAs. Science 294, 853.). Using HiPerFect Transfection Reagent, HeLa S3 cells (6 x 104 cells/well) were transfected with a luciferase reporter construct with a binding site for miR-1 in the 3' UTR; cotransfected with a luciferase reporter construct with a binding site for miR-1 in the 3' UTR and with a miR-1 Mimic; or transfected with a luciferase reporter construct with a binding site for miR-16 in the 3' UTR. Twenty-four hours later, luciferase activity was measured. miR-1 Mimic resulted in a similar decrease in luciferase level as endogenous miR-16.
1
miScript miRNA Mimic stable for 72 hours.
HeLa S3 cells (6 x 104 cells/well) were transfected with a negative control or an miR-1 Mimic using HiPerFect Transfection Reagent. Twenty-four hours prior to measurement, cells were transfected with a luciferase reporter construct carrying an miR-1 binding site using Attractene Transfection Reagent. After the time points indicated, miR-1 levels were detected by measuring luciferase activity.
2
Downregulation at protein level after miR-1 mimic transfection.
HDAC4 is predicted to be a target of miR-1 and has 2 miR-1 binding sites in the 3' UTR (Chen, J.F. et al. [2006] The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. Nat. Genet. 38, 228.). HeLa S3 cells (8 x 104 cells/well) were transfected using HiPerFect Transfection Reagent and a range of concentrations of miR-1 Mimic. miR-1 #1, miR-1 #2, and miR-1 #3 are different batches of miR-1 Mimic. After 24 and 48 hours, [A] HDAC4 protein level was analyzed by western blotting. Tubulin was also analyzed as an internal control (not shown). No effect was observed after 24 hours. After 48 hours, protein level was significantly decreased. [B] HDAC4 transcript level was analyzed by real-time PCR using the miScript System after transfection using 50 nM miR-1 Mimic. The negative control was AllStars Negative Control. Results were normalized to GAPDH and to the negative control and are presented as 2-ΔΔCT. Transcript levels did not change significantly at either the 24-hour or 48-hour time point.
Performance
miScript miRNA Mimics cause a decrease in gene expression similar to that observed with endogenous miRNAs (see figure "Comparable downregulation by endogenous miRNA and miScript miRNA Mimic"). The observed decrease in gene expression is often mediated at the translational level (see figure "Downregulation at protein level after miR-1 Mimic transfection"). miScript miRNA Mimics efficiently inhibit gene expression for up to 72 hours after transfection (see figure "miScript miRNA Mimic stable for 72 hours").
Principle

Transfection of mimics followed by downstream gene expression analysis or phenotypic analysis, is performed to elucidate the targets and roles of particular miRNAs. These experiments enable study of the biological effects of misregulation of individual miRNAs as well as confirmation of specific genes as targets of individual miRNAs. Reduced gene expression after transfection of mimic provides evidence that the miRNA under study is involved in regulation of that gene. miScript miRNA Inhibitors can also be used for similar studies. Typically, such experiments involve transfection of mimic or inhibitor, or alternatively cotransfection with a vector construct which carries the miRNA-binding site fused to a reporter gene. Downstream analysis can include reporter assays, real-time PCR, microarray analysis, or protein analysis.

Procedure

miScript miRNA Mimics are available for every human, mouse, rat, and virus miRNA in the current version of miRBase, the microRNA database. QIAGEN's GeneGlobe database is continually updated to ensure that mimic and inhibitor designs match the most up-to-date version of miRBase. This ensures that you are always working with the most accurate and current miRNA sequence information possible. If the miRNA of interest is not available in miRBase, a mimic for your human, mouse, rat, or virus miRNA can be custom synthesized. Simply visit the custom miScript miRNA product design page, enter a name for the mimic, provide the target sequence, and our design tool will provide a custom mimic ready to order.

Flexible formats enable either low-throughput analysis of small numbers of mimics or high-throughput screening of hundreds of mimics. miScript miRNA Mimics are available in tubes (1 nmol or 5 nmol) at cell-culture-grade (>90% purity provided by proprietary synthesis and purification processes). For in vivo applications, HPLC-purified, animal-grade miScript miRNA Mimics can be ordered at the 20 nmol scale (provided in tubes). For medium- to high-throughput applications, 96-well plates (1 nmol or 5 nmol), or 384-well plates (1 nmol) are available to suit your experimental requirements. Simply upload an Excel sheet of the predesigned (listed in miRBase) or custom (not listed in miRBase) miScript miRNA Mimics in your preferred plate layout, and then view and order the plate.

Applications
miRNA research
Gene regulation research
Features
Specifications
Design Predesigned and custom design avaliable
Format Single tubes, 96-well & 384 well plates
Modification None
Scale 1 nmol, 5 nmol, 20 nmol
Sequence provided No
Species Human, mouse, rat

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