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| 1. |
Kizaka-Kondoh, S., Itasaka, S., Zeng, L., Tanaka, S., Zhao, T., Takahashi, Y., Shibuya, K., Hirota, K., Semenza, G.L. and Hiraoka, M.
(2009)
Selective killing of hypoxia-inducible factor-1-actie cells improves survival in a mouse model of invasive and metastatic pancreatic cancer
Clin. Can. Res.
15
,
3433-3441
.
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Notes:
Hypoxia within solid tumors is a major factor that contributes to cancer progression. Pancreatic cancers express target genes of the transcription factor hypoxia-inducible factor-1. These genes encode proteins involved in angiogenesis, such as vascular endothelial growth factor (VEGF) and other growth factors like insulin-like growth factor (IGF) and proteins associated with the extracellular matrix. The authors of this study investigated whether HIF-1 activity plays a role in progression of pancreatic cancers. They implanted the human pancreatic cancer cells (SUIT-2) that had been transfected with a luciferase reporter in to nude male mice. Mice were either untreated or treated with the prodrug POP33, which appears to increase capsase-3 activity. They sought to determine if POP33 would induce apoptosis in HIF-1 expressing hypoxic cells. Bioluminescent imaging (BLI) was performed in vivo by injecting mice with D-luciferin solution. Caspase activity was also monitored in vivo using a luminescent caspase-3/7 substrate.
(0003997) |
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Products: VivoGlo™ Caspase-3/7 Substrate (Z-DEVD-Aminoluciferin) |
| 2. |
Xu, X., Soutto, M., Xie, Q., Servick, S., Subramanian, C., von Arnim, A.G., Johnson, C.H.
(2007)
Imaging protein interactions with bioluminescence resonance energy transfer (BRET) in plant and mammalian cells and tissues.
Proc. Natl. Acad. Sci. U S A
104
,
10264-10269
.
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Notes:
These authors used the coelenterazine analog ViviRen® as a substrate for Renilla luciferase in Bioluminescence Resonance Energy Transfer (BRET) experiments. Using this technique, they were able to demonstrate protein interactions in single mammalian cells.
(0003631) |
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Products: ViviRen™ Live Cell Substrate |
| 3. |
Dong, M., How, T., Kirkbride, K.C., Gordon, K.J., Lee, J.D., Hempel, N., Kelly, P., Moeller, B.J., Marks, J.R., and Blobe, G.C.
(2007)
The type III TGF-b receptor suppresses breast cancer progression.
J. Clin. Invest.
117
,
206-17
.
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Notes:
These authors showed that loss of the type II TGF-β receptor TGFβIII through allelic imbalance occurs during breast cancer development and increases metastatic potential. When TGFβIII expression was restored in human breast cancer cells, invasiveness, angiogenesis and metastasis were inhibited in an in vivo model system. The authors first analyzed TGFβIII mRNA levels using a cDNA array of 50 different breast cancer samples and controls. They also investigated TGFβIII protein expression levels by immunohistochemical analysis of a breast cancer tissue array containing over 250 breast cancers specimens at different stages of disease progression. Results from both analyses showed that TGFβIII expression decreased as disease progressed. The effect pf TGFβIII expression on tumor growth was investigated in a mouse model system. Murine 4T1 mammary cancer cells genetically engineered to express firefly luciferase were stably transfected with an expression vector containing TGFβIII , or a control vector. Tumor progression was then monitored in vivo by bioluminescent imaging. Cells expressing TGFβIII had delayed onset of metastasis and a reduction on the size and number of metastases compared with non-TGFβIII-expressing cells.
(0003617) |
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Products: Luciferin-EF™ |
| 4. |
Zhang, Y., So, M.-K., Loening, A.M., Yao, H., Gambhir, S.S., and Rao, J.
(2006)
HaloTag protein-mediated site-specific conjugation of bioluminescent proteins to quantum dots.
Angewandte Chemie International Edition
45
,
4536-4940
.
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Notes:
This paper describes work to use the HaloTag™ Interchangeable Protein Labeling Technology for the specific conjugation of proteins to quantum dots. The authors used the pHT2 vector to create a Renilla luciferase (Luc 8)/HaloTag™ Protein fusion that contained a 6X polyhistidine tag. The fusion protein was purified over a nickel affinity resin. They synthesized a HaloTag™ ligand for conjugation to the quantum dots. The ligand was designed to orient the ligand away from the quantum dot surface to minimize steric hindrance between the HaloTag™ fusion protein and the quantum dots upon interaction of the fusion protein with the ligand. The authors used BRET emission to evaluate conjugation of the HaloTag™ fusion protein to the quantum dots. They conclude that using this technology, they were able to conjugate a bioluminescent protein to quantum dots, creating self-illuminating quantum dots. Furthermore, they suggest that the mild conjugation conditions used may allow in vivo labeling of proteins or cells using quantum dots.
(0003490) |
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Products: HaloTag® pHT2 Vector |
| 5. |
Harmache, A., LeBerre, M., Droineau, S., Giovannini, M., and Bremont, M.
(2006)
Bioluminescence imaging of live infected Salmonids reveals that the fin bases are the major portal of entry for Novirhabdovirus.
J. Virol.
80
,
3655–3659
.
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Notes:
Researchers created a recombinant infectious hematopoietic necrosis virus (IHNVLUC) that contained the Renilla luciferase gene. The IHNVLUC construct was tested for its ability to infect and kill rainbow trout compared to other IHNV constructs. In other studies, the EnduRen™ Live Cell Substrate was used to monitor points of infection in live fish. It was found that Renilla luciferase could be detected within four days after trout were exposed to 5 x 104 PFU/ml of the IHNVLUC construct. Areas behind the fins were demonstrated to points of viral entry and infection.
(0003385) |
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Products: EnduRen™ Live Cell Substrate |
| 6. |
Zhang, W., Chen, M., West, D.B. and Purchio, A.F.
(2005)
Visualizing Drug Efficacy In Vivo
Molecular Imaging
4
,
88-90
.
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Notes:
The authors of this paper present proof-of-concept experiments showing that drug metabolism enzyme activity can be measured in whole animals (in vivo) in real time. Using a mouse that expresses a luciferase transgene at constitutively high levels in the liver, the authors evaluated CYP3A4 and CYP3A7 activity using a CYP3A P450 substrate (proluciferin substrate) that is converted into a luciferase substrate by CYP34 activity. The luciferase substrate produced by the P450 activity is then used by luciferase in a reaction that produces light. An increase in luminescence correlates with an increase in enzyme activity in this assay. The authors conclude that optical imaging of reporter mice will provide a new method for looking at drug actions in whole animals, with the caveat that the solubility of the proluciferin substrate is optimized and toxicity is minimized.
(0003996) |
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Products: P450-Glo™ CYP3A4 Assay | P450-Glo™ CYP3A7 Assay |
| 7. |
Grover, G.S., Turner, B.A., Parker, C.N., Meier, J., Lala, D. S., and Lee, P.H.
(2003)
Multiplexing Nuclear Receptors for Agonist Identification in a Cell-Based Reporter Gene High-Throughput Screen.
J. Biomol. Screen.
8
,
239-246
.
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Notes:
Nuclear receptor ligand binding regions from the Farnesoid-X-activated receptor (FXR) and peroxisome proliferator-activated receptor delta (PPARδ) were fused to a Gal4 DNA Binding Domain. These constructs were used to screen compounds from a LOPAC library of 640 compounds. Ligands bound to the Nuclear receptor ligand binding regions would result in the fusion proteins binding and inducing expression of a luciferase reporter construct with 5 upstream Gal4 binding element repeat sequences. For these experiments, Human hepatoma cell line (Huh7) cells transiently transfected with both plasmids were seeded in 384 well plates. After exposure to the various compounds in the LOPAC library luciferase expression was assessed with the Steady-Glo® Luciferase Assay System. Cells and Steady-Glo® reagent were added to cultures by a Labsystem Multidrop-384 liquid dispenser. Plates were read on a Molecular Devices ChemiLuminescence Imaging Plate Reader.
(0003265) |
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Products: Steady-Glo® Luciferase Assay System |
| 8. |
Yamamoto, Y.Y., Tsuhara, Y., Gohda, K., Suzuki, K. and Matsui, M.
(2003)
Gene trapping of the Arabidopsis genome with a firefly luciferase reporter.
Plant J.
35(2)
,
273-283
.
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Notes:
These researchers used luciferase-containing T-DNA insertions in Arabidopsis thaliana for gene trapping. Luciferase was chosen because its transient expresion allowed temporal expression studies. Several insertion vectors were constructed and found to have different insertion frequencies. Vectors containing the luc+ gene had substantially higher insertion rates than native luciferase vectors. Luciferase activity was measured in vivo with a CCD camera or, for longer term studies, with an automated scintillation counter sampling every 15-25 minutes over one week. The application of IRES sites in gene trapping experiments was also investigated using firefly and Renilla luciferases. The Dual-Luciferase® Reporter Assay System was used to monitor luciferase activity in vitro. Finally, to sequence the T-DNA insertion sites, genomic DNA was isolated from T2 seedlings using the Wizard® Magnetic 96 DNA Plant System and was subsequently amplified and sequenced.
(0002787) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pGEM®-luc DNA | pRL-TK Vector | pSP-luc+NF Fusion Vector | Wizard® Magnetic 96 DNA Plant System |