miRNA & 3’ UTR荧光素酶报告基因载体
genome-wide collection of pre-cloned luciferase 3´UTR reporters
The LightSwitch™ collection includes over 12,000 human endogenous 3`UTRs and hundreds of miRNA target bionsensors cloned into LightSwitch reporter vectors. These transfection-ready LightSwitch GoClones® together with the highly optimized assay reagents, transfection reagents, controls, miRNA Mimics and Inhibitors and detailed protocols allows you to perform your gene regulation reporter assay experiments immediately.
To ensure high-quality results, LightSwitch reporter vectors utilize a novel, engineered luciferase gene (RenSP) and optimized reagents that provide industry-leading sensitivity. For more information, please see the tabs below as well as the Overview and RenSP Luciferase sections on the Functional Genomics - LightSwitch™ page.
Click the button below to search the LightSwitch Reporter Collections and place your order on our affiliate website, SwitchGear Genomics.
Find and order LightSwitch™ GoClones®
To find LightSwitch GoClone® constructs that contain the promoters of your genes of interest, use the box below to search the LightSwitch Promoter Reporter Collection. Place your order using the specific Product ID(s) you find for the clone(s) you wish to order. As we cannot stock every GoClone® in all of our offices, delivery may take 10-14 days. Click here for Search Tips »
IMPORTANT: Because all LightSwitch reporter constructs contain the optimized RenSP luciferase gene, you MUST use our LightSwitch Luciferase Assay Reagents to obtain optimal results. (Other luciferase assay reagents are not compatible with RenSP.) We also recommend that you include appropriate positive & negative control vectors when you perform your assays.
Post transcriptional regulation through miRNA-3´UTR interactions
Gene regulation studies have historically focused on events that influence transcription, such as the binding of transcription factors to promoters. More recently, much research has centered on the presence or absence of histone modifications that alter the chromatin structure, which influences DNA accessibility and its ability to be transcribed. However, post-transcriptional events play an equally important role in gene regulation.
MicroRNAs (miRNAs) are small, non-coding RNA molecules of approximately 22 nucleotides that play a large role as post-transcriptional regulators. miRNAs are thought to repress expression by targeting the 3´UTR (untranslated) region of mRNA transcripts. The interaction of a miRNA with a 3´UTR results in either increased degradation of the target transcript or inhibition of translation.
While the human genome is thought to encode only about 1000 miRNAs, they are believed to target ~60% of mRNA transcripts. A given miRNA may have multiple 3´UTR targets, and a given 3´UTR may be targeted by multiple miRNAs. Therefore, there is considerable interest in studying miRNA-3´UTR interactions.
LightSwitch products for the study of 3´UTR-miRNA interactions
LightSwitch 3´UTR Reporter constructs are ideal for performing miRNA target validation, as well as assessing the functional impact of miRNA-3´UTR interactions. In combination with our miRNA Mimics and Inhibitors, which are co-transfected with the 3´UTR Reporter constructs (Figure 1), you have everything needed to validate miRNA targets, measure RNA stability, translation efficiency and the functional impact of miRNAs on a gene-by-gene basis. We also offer over 900 optimized LightSwitch Synthetic miRNA Target Reporter constructs, which can provide increased response relative to the endogenous sequences cloned into the LightSwitch 3´UTR Reporter constructs.
Figure 1: Luciferase activity used for target validation of miRNAs on a specific human 3´UTR.
- Transfect a 3´UTR Reporter construct into your cell line.
- A constitutive promoter drives the production of a hybrid luciferase-human 3´UTR transcript.
- Total luciferase output depends on the effect of the 3´UTR on transcript stability and/or translational efficiency.
- A synthetic microRNA or microRNA inhibitor can be co-transfected with the 3´UTR reporter vector to study its impact.
The 3´UTR activity can be measured to quantitively characterize:
- The impact of the UTR on post-transcriptional gene regulation
- The impact of a miRNA or siRNA on the regulation of transcript stability or translation efficiency
- The targets of a miRNA or siRNA (Figure 2)
- The effect of sequence variants on UTR function
- The impact of miRNA inhibitors or stimuli
Figure 2: Predicted mir-122 targets were validated using constructs from the LightSwitch 3´UTR Reporter Collection.
- Understand the mechanism by which a gene is induced or repressed.
- Measure the impact of the 3´UTR on post-transcriptional gene regulation.
- Validate the targets of a miRNA or siRNA.
- Measure the effects of sequence variants on 3´UTR or miRNA function.
Both the LightSwitch 3´UTR Reporter constructs and the LightSwitch Synthetic miRNA Target Reporter constructs were designed to be co-transfected with our collections of LightSwitch miRNA Mimics and Inhibitors to study the effects of over-expression of a miRNA of interest, or to knock down the expression of endogenous human miRNAs in living cells.
The following publications cite the use of and/or provide additional information about the LightSwitch Luciferase Assay System:
3´UTRs & miRNAs
- Myocardial MiR-30 downregulation triggered by doxorubicin drives alterations in β-adrenergic signaling and enhances apoptosis Roca-Alonso et al. (2015) Cell Death and Disease
- Microenvironment-induced downregulation of miR-193b drives ovarian cancer metastasis Mitra et al. (2015) Oncogene
- EGF induces microRNAs that target suppressors of cell migration: miR-15b targets MTSS1 in breast cancer Kedmi et al. (2015) Science Signaling
- MicroRNA-31 Promotes Skin Wound Healing by Enhancing Keratinocyte Proliferation and Migration Li et al. (2015) Journal of Investigative Dermatology
- Identification of microRNA-mRNA dysregulations in paroxysmal atrial fibrillation Chiang et al. (2015) International Journal of Cardiology
- Metformin and Rapamycin Reduce Pancreatic Cancer Growth in Obese Prediabetic Mice by Distinct MicroRNA-Regulated Mechanisms Cifarelli et al. (2015) Diabetes
- Identification and Computational Analysis of Gene Regulatory Elements Taher et al. (2015) Cold Spring Harb Protocols
- miR-155 augments CD8+ T-cell antitumor activity in lymphoreplete hosts by enhancing responsiveness to homeostatic γc cytokines Ji et al. (2014) PNAS
- PLK1 Inhibition Down-regulates Polycomb Group Protein BMI1 via Modulation of the miR-200c/141 Cluster Dimri et al. (2014) Journal of Biological Chemistry
- Mir-660 is downregulated in lung cancer patients and its replacement inhibits lung tumorigenesis by targeting MDM2-p53 interaction Fortunato et al. (2014) Cell Death and Disease
- miR30a inhibits LOX expression and anaplastic thyroid cancer progression Boufraqech et al. (2014) Cancer Research
- MicroRNA silencing for cancer therapy targeted to the tumour microenvironment Cheng et al. (2014) Nature
- TRPM3 and miR-204 Establish a Regulatory Circuit that Controls Oncogenic Autophagy in Clear Cell Renal Cell Carcinoma Hall et al. (2014) Cancer Cell
- miR-24 limits aortic vascular inflammation and murine abdominal aneurysm development Maegdefessel et al. (2014) Nature Communications
- miR-154* and miR-379 in the DLK1-DIO3 MicroRNA Mega-Cluster Regulate Epithelial to Mesenchymal Transition and Bone Metastasis of Prostate Cancer Gururajan et al. (2014) Clinical Cancer Research
- MiR-29b replacement inhibits proteasomes and disrupts aggresome+autophagosome formation to enhance the antimyeloma benefit of bortezomib Jagannathan et al. (2014) Leukemia
- The Regulatory Function of miR-200c on Inflammatory and Cell-Cycle Associated Genes in SK-LMS-1, A Leiomyosarcoma Cell Line Chuang et al. (2014) Reproductive Sciences
- Integrative post-genome-wide association analysis of CDKN2A and TP53 SNPs and risk of esophageal adenocarcinoma Buas et al. (2014) Carcinogenesis
- Kaposi’s Sarcoma-Associated Herpesvirus Downregulates Transforming Growth Factor β2 To Promote Enhanced Stability of Capillary-Like Tube Formation DiMaio et al. (2014) Journal of Virology
- MicroRNA-193b-3p acts as a tumor suppressor by targeting the MYB oncogene in T-cell acute lymphoblastic leukemia Mets et al. (2014) Leukemia
- Human microRNA-24 modulates highly pathogenic avian-origin H5N1 influenza A virus infection in A549 cells by targeting secretory pathway furin Loveday et al. (2014) Journal of General Virology
- Renilla Luciferase Reporter Assay to Study 3´UTR-Driven Posttranscriptional Regulations of OPRM1 Vincelli et al. (2014) Methods in Molecular Biology
- microRNA100 inhibits self-renewal of breast cancer stem-like cells and breast tumor development Deng et al. (2014) Cancer Research
- Selective repression of the oncogene cyclin D1 by the tumor suppressor miR-206 in cancers Elliman et al. (2014) Oncogenesis
- Stromal fibroblast-derived miR-409 promotes epithelial-to-mesenchymal transition and prostate tumorigenesis Josson et al. (2014) Oncogene
- MicroRNA-138 is a potential regulator of memory performance in humans Schröder et al. (2014) Frontiers in Human Neuroscience
- miR-150 influences B-cell receptor signaling in chronic lymphocytic leukemia by regulating expression of GAB1 and FOXP1 Mraz et al. (2014) Blood
- MicroRNA-34a Modulates Neural Stem Cell Differentiation by Regulating Expression of Synaptic and Autophagic Proteins Morgado et al. (2014) Molecular Neurobiology
- Circulating microRNA signature in non-alcoholic fatty liver disease: from serum non-coding RNAs to liver histology and disease pathogenesis Pirola et al. (2014) Gut
- A Novel Function for Lysyl Oxidase in Pluripotent Mesenchymal Cell Proliferation and Relevance to Inflammation-Associated Osteopenia Khosravi et al. (2014) PLOS One
- Regulation of COX-2 expression by miR-146a in lung cancer cells Cornett et al. (2014) RNA
- MicroRNA-205 signaling regulates mammary stem cell fate and tumorigenesis Chao et al. (2014) Journal of Clinical Investigation
- CFIm25 links alternative polyadenylation to glioblastoma tumour suppression Masamha et al. (2014) Nature
- Therapeutic Delivery of miR-200c Enhances Radiosensitivity in Lung Cancer Cortez et al. (2014) Molecular Therapy
- miR-145 suppresses thyroid cancer growth and metastasis and targets AKT3 Boufraqech et al. (2014) Endocrine-Related Cancer
- Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode Agrawal et al. (2014) PNAS
- Assessment of microRNA-related SNP effects in the 3´ untranslated region of the IL22RA2 risk locus in multiple sclerosis Lill et al. (2014) Neurogenetics
- Long Non-coding RNA HOTAIR Is Targeted and Regulated by miR-141 in Human Cancer Cells Chiyomaru et al. (2014) Journal of Biological Chemistry
- Systematic design and functional analysis of artificial microRNAs Arroyo et al. (2014) Nucleic Acids Research
- Identifying microRNAs regulating B7-H3 in breast cancer: the clinical impact of microRNA-29c Nygren et al. (2014) British Journal of Cancer