通过合成miRNA进行miRNA功能和基因调控研究

  • 模拟内源性miRNAs,可直接用于转染
  • 基于miRBase中的所有人类、小鼠和大鼠的miRNAs而设计
  • 可灵活选择规格、包装和纯度
  • 为miRBase中未列出的miRNAs提供订制设计
  • 可通过GeneGlobe便利的订购
miScript miRNA Mimics是化学合成的双链RNAs,在转染到细胞后模拟内源成熟miRNAs。提供细胞培养级纯度(>90%纯度)和动物级纯度(经HPLC纯化;用于体内实验)的模拟物。如需进行灵活的筛选实验,可选择miScript miRNA Mimic Plates,以96孔板或384孔板形式提供预制或用户订制的miScript miRNA Mimics。miScript miRNA Inhibitors也可用于类似的实验。如需进行转染实验,QIAGEN提供HiPerFect Transfection Reagent进行miRNA Mimic转染实验,Attractene Transfection Reagent进行miRNA Mimic-DNA共转染实验。
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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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
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miScript miRNA Mimics are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


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Comparable downregulation by endogenous miRNA and miScript miRNA Mimic.|miScript miRNA Mimic stable for 72 hours.|Downregulation at protein level after miR-1 mimic transfection.|
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.|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.|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与内源性miRNAs相同会使得基因表达下降(参见"Comparable downregulation by endogenous miRNA and miScript miRNA Mimic")。通常在翻译水平观察到基因表达下降(参见"Downregulation at protein level after miR-1 Mimic transfection")。转染后,miScript miRNA Mimics可有效抑制基因表达至多72小时(参见"miScript miRNA Mimic stable for 72 hours")。
Principle

模拟物转染及下游基因表达分析或表型分析,用于确定特定miRNAs的靶向和作用。这些实验能够研究个体miRNAs失调的生物效应及确定作为个体miRNAs靶向的特异性基因。抑制剂转染后基因表达增强表明研究中的miRNA由该基因调控。类似的研究也可使用miScript miRNA Inhibitors。一般而言,这样的实验都要涉及到模拟物或抑制剂的转染或与携带miRNA结合位点融合报告基因的载体结构共转染。下游分析包括报告分析、real-time PCR、微阵列分析或蛋白质分析。

Procedure

miScript miRNA Mimics提供适合当前版本microRNA数据库(miRBase)中的人类、小鼠、大鼠和病毒miRNA。QIAGEN's GeneGlobe数据库持续更新数据,确保模拟物和抑制剂与最新的miRBase中数据尽量匹配。确保得到高度准确、当前的miRNA序列信息。若miRBase中没有您所需的miRNA的数据,可访问用户订制miScript miRNA产品设计页面,输入模拟物名称,提供目标序列,我们的设计工具即可提供您所需的模拟物,供您订购。

可灵活选择规格,用少量模拟物进行低通量分析或大量模拟物进行高通量筛查实验。miScript miRNA Mimics以试管形式(1 nmol或5 nmol)提供细胞培养级纯度(>90%,由专门的合成和纯化流程提供)的模拟物。用于体内实验,经HPLC纯化、动物级纯度的miScript miRNA Mimics也可订购得到,规格为20 nmol(以试管形式提供)。对于中到高通量应用,提供96孔板(1 nmol或5 nmol)或384孔板形式(1 nmol),满足您的实验需求。只需上传预设计的Excel表格(miRBase中列出)或订制(miRBase中未列出的)miScript miRNA Mimics,选择你所需的孔板形式,即可查询并订购孔板。

Applications
  • miRNA研究
  • 基因调控研究
Feature
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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Safety Data Sheets
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