Luciferase Assay Kits
GeneCopoeia offers genome-wide human, mouse and rat microRNA (miRNA) 3' UTR target clones in mammalian expression vectors. miRNA 3' UTR target clones can be used for miRNA target identification and functional validation of predicted targets or to study the regulatory effect of miRNA on target gene(s).
3' UTR sequences were obtained from public domain gene sequence databases and inserted downstream of the secreted Gaussia luciferase (GLuc) reporter gene in its respective vector system, driven by SV40 promoter for expression in mammalian cells. A chimeric mRNA is transcribed consisting of the GLuc and a 3' UTR target sequence. Thus, the study of mRNA-miRNA target interaction can be easily performed with a live cell assay for GLuc using only 10 μl of the cell culture medium.
Besides using GLuc as the miRNA 3' UTR target reporter, a secreted Alkaline Phosphatase (SEAP) reporter driven by a CMV promoter, is also cloned into the same vector (pEZX-MT05) and serves as the internal control. The dual-reporter vector system enables transfection-normalization for accurate across-sample comparison.
In addition to the GLuc/SEAP dual-reporter vector system, a firefly/Renilla Duo-Luciferase reporter vector (pEZX-MT01) is also available.
|Figure 1. How 3’UTR clone works|
|Figure 2. Vector backbone of miTarget miRNA 3' UTR target clones|
Live cell assays
- Naturally secreted GLuc reporter
- No lysis of the cells is necessary
- Save samples, reduce variations, and simplify experiments
Dual-reporter vector system
- Enables transfection-normalization for accurate across-sample comparison
- Group study or high sample number compatible
- GLuc is 1000-fold more sensitive than firefly or Renilla luciferase
- All 3' UTR target clones are transfection-ready
3' UTR target vectors and compatible assay kits
|Vector||Reporter gene||Tracking gene||Advantage||Compatible assay kit|
|pEZX-MT05||Gaussia luciferase||Secreted alkaline phosphatase (SEAP)||Live cell assays||Secrete Pair ™ Dual Luminiscence Assay Kit|
|pEZX-MT01||Firefly luciferase||Renilla luciferase||Cell lysis required||Luc-Pair™ Duo-Luciferase Assay Kit|
|For miRNA target validation using GeneCopoeia's pEZX-MT05 and pEZX-MT01 target clones, our Secrete-Pair™ Dual Luminescence Assay Kit and Luc-Pair™ Duo-Luciferase Assay Kit are the recommended assay kits respectively. Click here for more information regarding validation of pEZX-MT01 clones.|
Pre-designed 3' UTR target clones
Search over 18,000 human, 18,000 mouse, and 10,000 rat pre-designed 3' UTR target clones in your choice of vector.
|MiTarget™ MicroRNA 3' UTR Target Clone Cancellation, QA, License, and Warranty|
- Kulyté, A. et al (2013) Additive effects of miRNAs and transcription factors on CCL2 production in human white adipose tissue. Diabetes, Published online before print December 30, 2013, doi: 10.2337/db13-0702 . [3'UTR of SP1].
- Sturrock, A. et al (2013) Key role of microRNA in the regulation of Granulocyte-macrophage colony stimulating factor expression in murine alveolar epithelial cells during oxidative stress. The Journal of Biological Chemistry, First Published on December 26, 2013, doi: 10.1074/jbc.M113.535922 jbc.M113.535922.. [dual- reporter expression clone of murine GM-CSF 3' UTR].
- Xu, S. et al (2013) Micrornas as new therapeutic targets for the prevention and/or treatment of retinopathy. United States Patent Application, US20130331436 A1. [Luciferase target reporter vector, pEZX-MT01].
- Ubhi, K. et al (2013) Widespread microRNA dysregulation in multiple system atrophy–disease‐related alteration in miR‐96. European Journal of Neuroscience, first published online: 5 DEC 2013 DOI: 10.1111/ejn.12444. [Reporter clones].
- Loyer, X. et al (2013) Inhibition of microRNA-92a Prevents Endothelial Dysfunction and Atherosclerosis in Mice. Circulation Research, doi: 10.1161/CIRCRESAHA.114.302213. [3' UTR target clone of SOCS5].
- Liang, Y. et al (2013) Requirement of miR‐144 in CsA Induced Proliferation and Invasion of Human Trophoblast Cells by Targeting Titin. Journal of Cellular Biochemistry, DOI: 10.1002/jcb.24710. [3' UTR target clone].
- Feng, Y. et al (2013) Heat shock improves sca‐1+ stem cells survival and directs ischemic cardiomyocytes towards a prosurvival phenotype via exosomal transfer: a critical role for HSF1/ …. STEM CELLS, Volume 32, Issue 2, pages 462–472, February 2014. [pEZX-MT01-HSP70-3'UTR target clone].
- Ratovitski, EA. et al (2013) Phospho-ΔNp63α regulates AQP3, ALOX12B, CASP14 and CLDN1 expression through transcription and microRNA modulation. FEBS letters, Volume 587, Issue 21, 1 November 2013, Pages 3581–3586. [HmiT006376-MT01].
- Robinson, PM. et al (2013) MicroRNA Signature in Wound Healing Following Excimer Laser Ablation: Role of miR-133b on TGFβ1, CTGF, SMA, and COL1A1 Expression Levels in Rabbit …. Investigative Ophthalmology & visual science, October 23, 2013 vol. 54 no. 10 6944-6951. [3' UTR target clones of CTGF and TGFβ1].
- Yu, B. et al (2013) Enhanced mesenchymal stem cell survival induced by GATA-4 overexpression is partially mediated by regulation of the miR-15 family. The International Journal of Biochemistry & Cell Biology, Volume 45, Issue 12, December 2013, Pages 2724–2735. [3'UTR OF Bcl-2l2 (Bcl-w)].
- Lindenbergh-van, Marlon. et al (2013) Identification of lethal microRNAs specific for head and neck cancer. Clinical Cancer Research, Clin Cancer Res October 15, 201319; 5647. [Luciferase reporter construct containing the 3'UTR sequence of ATM].
- Shukla, U. et al (2013) Insights into insulin-mediated regulation of CYP2E1: miR 132/212 targeting of CYP2E1 and role of PI3-K, Akt, mTOR signaling in regulating miR132/212 and miR 122/ …. Drug Metabolism & Disposition, doi:10.1124/dmd.113.052860. [Scrambled control expression vector (CmiR0001-MR04), Luc-Pair miR luciferase assay ].
- Kubo, Eri. et al (2013) Dynamic and differential regulation in the microRNA expression in the developing and mature cataractous rat lens. Journal of Cellular and Molecular Medicine, Volume 17, Issue 9, pages 1146–1159, September 2013. [TM2β 3'-UTR miRNA Target clone].
- Lin, HY. et al (2013) STAT3 upregulates miR-92a to inhibit RECK expression and to promote invasiveness of lung cancer cells. British Journal of Cancer, (2013) 109, 731–738.. [Control reporter and 3'-UTR reporter plasmid of RECK gene].
- Bier, A. et al (2013) MicroRNA-137 is downregulated in glioblastoma and inhibits the stemness of glioma stem cells by targeting RTVP-1. Oncotarget, Oncotarget. 2013 May; 4(5): 665–676. [plasmids expressing RTVP-1 and luciferase-RTVP-1-3'UTR].
- Ling, S . et al (2013) Dickkopf-1 (DKK1) phosphatase and tensin homolog on chromosome 10 (PTEN) crosstalk via microRNA interference in the diabetic heart. Basic Research in Cardiology, May 2013, 108:352. [Luc-PairTM miR Luciferase Assay Kit].
- Qu, Y. et al (2013) MiR‐182 and miR‐203 induce mesenchymal to epithelial transition and self‐sufficiency of growth signals via repressing SNAI2 in prostate cells. International Journal of Cancer, DOI: 10.1002/ijc.28056. [3′-UTR of SNAI2 in the pEZX-MT01 vector]
- Sundaram, G. et al (2013) /See-saw/'expression of microRNA-198 and FSTL1 from a single transcript in wound healing. Nature, 495, 103-106. doi:10.1038/nature11890 [3′UTR clone].
- Schrum, L. et al (2013) Treatment of Fibrosis Using Microrna 19b. U.S. Patent No. 20,130,053,429. 28 Feb. 2013. [3′UTR of TGFβRII in pEZX-MT01].
- Chiang, C. et al (2013) Up-regulation of miR-182 by β-catenin in breast cancer increases tumorigenicity and invasiveness by targeting the matrix metalloproteinase inhibitor RECK. Biochimica et Biophysica Acta, Volume 1830, Issue 4, April 2013, Pages 3067–3076. [3′UTR of RECK in pEZX-MT01].
- Wang, J. et al (2013) MicroRNA-31 controls phenotypic modulation of human vascular smooth muscle cells by regulating its target gene cellular repressor of E1A-stimulated genes. Experimental Cell Research, online 19 March 2013. [3’UTR of CREG in pEZX-MT01].
- Sengupta, J. et al (2013) MicroRNA-mediated GABA receptor subunit down-regulation in adult spinal cord following neonatal cystitis-induced chronic visceral pain in rats. PAIN, Volume 154, Issue 1, January 2013, Pages 59–70. [3′UTR of GABA Aα-1 gene in pEZX-MT01].
- Zhang, L. et al (2013) MiR-206 inhibits gastric cancer proliferation in part by repressing CyclinD2. Cancer Letters, Volume 332, Issue 1, 10 May 2013, Pages 94–101. [3′-UTR of CCND2 with miR-206].
- Dakhlallah, D. et al (2013) Epigenetic regulation of miR-17∼ 92 contributes to the pathogenesis of pulmonary fibrosis. American Journal of Respiratory and Critical Care Medicine, 187.4 (2013): 397-405. [3′ UTR of Col13a in pEZX-MT05 reporter vector].
- Devany, E. et al (2013) Positive and negative feedback loops in the p53 and mRNA 3′ processing pathways. Proceedings of the National Academy of Sciences, 110.9 (2013): 3351-3356. [Luciferase vector pEZX-MT01 with TP53 microRNA 3′ UTR target clone; catalog# HmiT054283-MT01].
- Majid, S. et al (2013) miRNA-34b Inhibits Prostate Cancer through Demethylation, Active Chromatin Modifications, and AKT Pathways. Clin Cancer Res January 1, 2013 19; 73 doi: 10.1158/1078-0432.CCR-12-2952. [3′-UTR target expression clones].
- Jin, Y. et al (2013) Evaluating the MicroRNA Targeting Sites by Luciferase Reporter Gene Assay. MicroRNA Protocols, Humana Press, 2013. 117-127. [luciferase constructs containing entire 3′-UTR].
- Makino, K. et al (2013) The Downregulation of microRNA let-7a Contributes to the Excessive Expression of Type I Collagen in Systemic and Localized Scleroderma. Journal of Immunology, doi: 10.4049/jimmunol.1200822. [luciferase construct containing a2(I) collagen 39-UTR].
- Rolland-Turner, M. et al (2013) Adenosine Stimulates the Migration of Human Endothelial Progenitor Cells. Role ofCXCR4 and MicroRNA-150. PloS one, 8(1): e54135. doi:10.1371/journal.pone.0054135. [CXCR4 3’UTR in pEZX-MT05].
- Goretti, E. et al (2013) MicroRNA-16 affects key functions of human endothelial progenitor cells. Journal of Leukocyte Biology, doi: 10.1189/jlb.1012511 jlb.1012511. [CCND1 3’UTR].
- Klein, D. et al (2013) MicroRNA Expression in Alpha and Beta Cells of Human Pancreatic Islets. PloS one, 8(1): e55064. doi:10.1371/journal.pone.0055064. [cMaf 3′UTR in pEZX-MT01].
- Lee, H.K. et al (2013) MicroRNA-145 Is Downregulated in Glial Tumors and Regulates Glioma Cell Migration by Targeting Connective Tissue Growth Factor. PloS one, 8(2): e54652. doi:10.1371/journal.pone.0054652. [luciferase-CTGF-3′UTR].
- Rosario, R. et al. (2013) Comparative study of microRNA regulation on< i> FOXL2</i> between adult-type and juvenile-type granulosa cell tumours< i> in vitro</i>. Gynecologic Oncology, Volume 129, Issue 1, April 2013, Pages 209–215. [3’UTR target clone for FOXL2 (HmiT017665-MT01)].
- Zehavi, L. et al (2012) Silencing of a large microRNA cluster on human chromosome 14q32 in melanoma: biological effects of mir-376a and mir-376c on insulin growth factor 1. Molecular Cancer 2012, 11:44. [3’UTR target clone for IGF1R (HmiT009523-MT01)].
- Muratsu-Ikeda, S. et al. (2012) Downregulation of miR-205 Modulates Cell Susceptibility to Oxidative and Endoplasmic Reticulum Stresses in Renal Tubular Cells. PLoS ONE 7(7): e41462. doi:10.1371/journal.pone.0041462. [EGLN2 3′UTR miRNA target clone].
- Amodio, N. et al. (2012) DNA-demethylating and anti-tumor activity of synthetic miR-29b mimics in multiple myeloma. Oncotarget, 3.10 (2012): 1246-1258. [3’UTRs of DNMT3a and DNMT3B].
- Fang, Y. et al. (2012) Site-Specific MicroRNA-92a Regulation of Krüppel-Like Factors 4 and 2 in Atherosusceptible Endothelium. Arteriosclerosis, Thrombosis, and Vascular Biology. 2012; 32: 979-987 [3’UTR of human KLF4 and KLF2].
- Sonkoly, E. et al. (2012) MicroRNA-203 functions as a tumor suppressor in basal cell carcinoma. Oncogenesis (2012) 1, e3; doi:10.1038/oncsis.2012.3. [3’UTR of c-JUN].
- Alla, V. et al. (2012) E2F1 confers anticancer drug resistance by targeting ABC transporter family members and Bcl-2 via the p73/DNp73-miR-205 circuitry. Cell Cycle, Volume 11, Issue 16 August 15, 2012. [3’UTR of E2F1].
- Lai, V. et al. (2012) MicroRNA-143 is a critical regulator of cell cycle activity in stem cells with co-overexpression of Akt and angiopoietin-1 via transcriptional regulation of Erk5/cyclin D1 signaling. Cell Cycle, Volume 11, Issue 4 February 15, 2012. [miR-143, 3’UTR of Erk5].
- Yang, Y. et al. (2012) Decrease of miR-122 causes hepatic insulin resistance by inducing protein tyrosine phosphatase 1B, which is reversed by licorice flavonoid. Hepatology, DOI: 10.1002/hep.25912. [3’UTR of PTP1B].
- Angel-Morales, G. et al. (2012) Red wine polyphenolics reduce the expression of inflammation markers in human colon-derived CCD-18Co myofibroblast cells: Potential role of microRNA-126. Food Funct., 2012,3, 745-752. [3’UTR of VCAM-1].
- Zehavi, L. et al. (2012) Silencing of a large microRNA cluster on humanchromosome 14q32 in melanoma: biological effects of mir-376a and mir-376c on insulin growth factor 1 receptor. Molecular Cancer 2012, 11:44. [3’UTR of IGF1R].
- Couts, K. et al. (2012) Oncogenic B-Raf signaling in melanoma cells controls a network of microRNAs with combinatorial functions. Oncogene advance online publication 2 July 2012; doi: 10.1038/onc.2012.209. [3’UTR].
- Yin, K. et al. (2012) Vascular endothelial cell-specific microRNA-15a inhibits angiogenesis in hindlimbischemia. The Journal of Biological Chemistry, 287, 27055-27064. [3′UTR of human VEGFa].
- Tanic, M. et al. (2012) Integration of BRCA1-mediated miRNA and mRNA profiles reveals microRNA regulation of TRAF2 and NFκBpathway. Breast Cancer Research and Treatment Volume 134, Number 1 (2012), 41-51, DOI: 10.1007/s10549-011-1905-4. [3’UTR of TRAF2].
- Arner, E. et al. (2012) Adipose tissue microRNAs as regulators of CCL2 production in human obesity. Diabetes August 2012 vol. 61 no. 8 1986-1993. [3′UTR of CCL2, ARNT, MAX, and RELB].
- Palumbo, T. et al. (2012) Functional screen analysis reveals miR-26b and miR-128 as central regulators of pituitary somatomammotrophic tumor growth through activation of the PTEN–AKT pathway. Oncogene, (21 May 2012) | doi:10.1038/onc.2012.190. [3’UTR of PTEN, BMI1].
- Chuang, T. et al. (2012) miR-200c is aberrantly expressed in leiomyomas in an ethnic-dependent manner and targets ZEBs, VEGFA, TIMP2, and FBLN5. Endocr Relat Cancer August 1, 2012 19 541-556. [3′ UTR of ZEB1, ZEB2, VEGFA, TIMP2, FBLN5].
- Ibrahim, S. et al. (2012) Targeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motility and invasiveness via a Rho-GTPase-and E-cadherin-dependent mechanism. International Journal of Cancer, Volume 131, Issue 6, pages E884–E896, 15 September 2012. [3’UTR of human SDC1].
- Zhuang, G. et al. (2012) Tumour-secreted miR-9 promotes endothelial cell migration and angiogenesis by activating the JAK-STAT pathway. The EMBO Journal (2012) 31, 3513 – 3523 doi:10.1038/emboj.2012.183. [3′UTR of human SOCS5].
- Lee, CG. et al. (2012) Farnesoid X receptor protects hepatocytes from injury by repressing miR-199a-3p, which increases levels of lkb1. Gastroenterology, Volume 142, Issue 5, May 2012, Pages 1206–1217.e7. [3′UTR of LKB1].
- Vickers, K. et al. (2012) MicroRNA-27b is a regulatory hub in lipid metabolism and is altered in dyslipidemia. Hepatology online: 6 JUL 2012 , DOI: 10.1002/hep.25846. [3’UTR of human GPAM].
- Li, Y. et al. (2012) Epigenetic deregulation of miR-29a and miR-1256 by isoflavone contributes to the inhibition of prostate cancer cell growth and invasion. Epigenetics Volume 7, Issue 8 August 2012. [TRIM68 3′UTR].
- Bose, D. et al. (2012) The Tuberculosis Drug Streptomycin as a Potential Cancer Therapeutic: Inhibition of miR-21 Function by Directly Targeting Its Precursor. Angewandte Chemie, Volume 124, Issue 4, pages 1043–1047, January 23, 2012. [3’UTR clone].
- Yuan, K. et al. (2012). Role of miR-148a in Hepatitis B Associated Hepatocellular Carcinoma. PLoS ONE 7(4): e35331. doi:10.1371/journal.pone.0035331. [3′UTR of miR-148a].
- Saini, S. et al. (2012) miRNA-708 Control of CD44+ Prostate Cancer–Initiating Cells. Cancer Res; 72(14); 3618–30. [3’UTR of CD44 and AKT2].
- Biyashev, et al. (2012) miR-27b controls venous specification and tip cell fate. Blood March 15, 2012 vol. 119 no. 11 2679-2687. [3’UTR of Spry2].
- Panda, H. et al. (2012) Endometrial miR-200c is altered during transformation into cancerous states and targets the expression of ZEBs, VEGFA, FLT1, IKKβ, KLF9, and FBLN5. Reproductive Sciences August 2012 vol. 19 no. 8 786-796. [3′UTR of ZEB1, ZEB2, VEGFA, FBLN5, KLF9].
- Panda, H. et al. (2012) Endometrial miR-181a and miR-98 Expression Is Altered during Transition from Normal into Cancerous State and Target PGR, PGRMC1, CYP19A1, DDX3X, and TIMP3. The Journal of Clinical Endocrinology & Metabolism July 1, 2012 vol. 97 no. 7 E1316-E1326. [3’UTR of PGRMC1 and DDX3X].
- Mackiewicz, M. et al. (2012) Identification of the receptor tyrosine kinase AXL in breast cancer as a target for the human miR-34a microRNA. Breast Cancer Research and Treatment Volume 130, Number 2 (2011), 663-679, DOI: 10.1007/s10549-011-1690-0. [3’UTR of AXL].
- Dippold, R. et al. (2012) Chronic ethanol feeding enhances miR-21 induction during liver regeneration while inhibiting proliferation in rats. AJP – GI July 12, 2012. [3’UTR].