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#33264648   2020/11/29 To Up

Development of a CRISPR/Cas9 system in Entamoeba histolytica: proof of concept.

The protozoan parasite Entamoeba histolytica is an important human pathogen and a leading parasitic cause of death on a global scale. The lack of molecular tools for genome editing hinders the study of important biological functions of this parasite. Due to its versatility, the CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 system has been successfully used to induce site-specific genomic alterations, including in protozoan parasites. In this study, we optimised CRISPR-Cas9 for use as a genetic tool in E. histolytica. We chose a single plasmid approach containing both guide RNA (gRNA) and Cas9 nuclease expression cassettes. The amebic U6 promoter was used to drive the expression of the gRNA and its expression was confirmed by Northern blot analysis. Stable transfectant cell lines were obtained using a destabilising domain of dihydrofolate reductase fused to myc-tagged Cas9 (ddCas9). With this system, we were able to induce ddCas9 expression 16 h following treatment with the small molecule ligand trimethoprim (TMP). Stable cell lines expressing ddCas9 and Luc-gRNA or non-specific (NS)-gRNA were transiently transfected with a plasmid containing a mutated luciferase gene (pDeadLuc) targeted by Luc-gRNA and another plasmid with a truncated luciferase gene (pDonorLuc) to restore luciferase expression and consequent activity. We observed that luminescence signal increased for the cell line expressing Luc-gRNA, suggesting that homologous recombination was facilitated by Cas9 activity. This evidence is supported by the presence of chimeric DNA detected by PCR and confirmed by sequencing of the resulting repaired DNA obtained by homologous recombination. We believe this represents the first report of a CRISPR/Cas9 system use in Entamoeba and provides evidence that this genome editing approach can be useful for genetic studies in this early branching eukaryote.
Monica Mendes Kangussu-Marcolino, Pedro Morgado, Dipak Manna, Heather Yee, Upinder Singh

2677 related Products with: Development of a CRISPR/Cas9 system in Entamoeba histolytica: proof of concept.

100 0.1ml (1mg/ml)1 mL300 units96 assays 70 Slides

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#32698955   2020/07/19 To Up

Dual targeting of GSK3B and HDACs reduces tumor growth and improves survival in an ovarian cancer mouse model.

To investigate the anti-tumor effect of a newly-developed dual inhibitor (APCS-540) of glycogen synthase kinase 3 beta (GSK3B) and histone deacetylases (HDACs) in ovarian cancer cells.
Enes Taylan, Fouzia Zayou, Ramachandran Murali, Beth Y Karlan, Stephen J Pandol, Mouad Edderkaoui, Sandra Orsulic

1871 related Products with: Dual targeting of GSK3B and HDACs reduces tumor growth and improves survival in an ovarian cancer mouse model.

100.00 ug100ug100.00 ug100ug100.00 ug100.00 ug100.00 ug100.00 ug0.1ml (1mg/ml)

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#32171747   2020/06/24 To Up

ABL1, Overexpressed in Hepatocellular Carcinomas, Regulates Expression of NOTCH1 and Promotes Development of Liver Tumors in Mice.

We investigated whether ABL proto-oncogene 1, non-receptor tyrosine kinase (ABL1) is involved in development of hepatocellular carcinoma (HCC).
Fang Wang, Wei Hou, Lennox Chitsike, Yingchen Xu, Carlee Bettler, Aldeb Perera, Thomas Bank, Scott J Cotler, Asha Dhanarajan, Mitchell F Denning, Xianzhong Ding, Peter Breslin, Wenan Qiang, Jun Li, Anthony J Koleske, Wei Qiu

2157 related Products with: ABL1, Overexpressed in Hepatocellular Carcinomas, Regulates Expression of NOTCH1 and Promotes Development of Liver Tumors in Mice.

300 units

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#32009285   2020/03/10 To Up

An eggplant SmICE1a gene encoding MYC-type ICE1-like transcription factor enhances freezing tolerance in transgenic Arabidopsis thaliana.

Low temperature is a crucial environmental factor affecting the quality and production of eggplant. Therefore, it is necessary to explore the molecular mechanisms of low temperature response. We isolated an ICE (inducer of CBF expression) gene from Solanum melongena, named SmICE1a. We then analysed structure, transcriptional activity and expression patterns of SmICE1a. Moreover, we also expressed SmICE1a in Arabidopsis thaliana. Bioinformatics and expression analysis showed that SmICE1a has a typical S-rich motif, ZIP region, bHLH and ACT-like domain. The gene SmICE1a had transcriptional activity in yeast and was localized to the nucleus following transient expression in tobacco leaves, which suggests that SmICE1a is a transcription factor. A dual-LUC assay revealed that SmICE1a can enhance expression of SmCBF. Overexpression of SmICE1a in Arabidopsis increased freezing tolerance and caused multiple biochemical changes: transgenic lines have higher proline content and lower electrolyte leakage and malondialdehyde than the wild type in cold conditions. The expression of AtCBF and their target genes, AtCOR15A, AtCOR47, AtKIN1 and AtRD29A, were up-regulated in SmICE1a-overexpressing plants under low temperatures. Based on these results, we suggest that SmICE1a plays an important role in cold response, which may help to understand the cold response mechanism in eggplant and could be used to enhance cold tolerance of eggplant in future.
L Zhou, Y J He, J Li, L Z Li, Y Liu, H Y Chen

2526 related Products with: An eggplant SmICE1a gene encoding MYC-type ICE1-like transcription factor enhances freezing tolerance in transgenic Arabidopsis thaliana.

25UG100 ug100.00 ug1 mL20 100 μg100ul4 Membranes/Box100.00 ug20 100.00 ug100 μg

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#31189617   2019/06/12 To Up

Diagnosis of obstructive coronary artery disease using computed tomography angiography in patients with stable chest pain depending on clinical probability and in clinically important subgroups: meta-analysis of individual patient data.

To determine whether coronary computed tomography angiography (CTA) should be performed in patients with any clinical probability of coronary artery disease (CAD), and whether the diagnostic performance differs between subgroups of patients.
Robert Haase, Peter Schlattmann, Pascal Gueret, Daniele Andreini, Gianluca Pontone, Hatem Alkadhi, Jörg Hausleiter, Mario J Garcia, Sebastian Leschka, Willem B Meijboom, Elke Zimmermann, Bernhard Gerber, U Joseph Schoepf, Abbas A Shabestari, Bjarne L Nørgaard, Matthijs F L Meijs, Akira Sato, Kristian A Ovrehus, Axel C P Diederichsen, Shona M M Jenkins, Juhani Knuuti, Ashraf Hamdan, Bjørn A Halvorsen, Vladimir Mendoza-Rodriguez, Carlos E Rochitte, Johannes Rixe, Yung Liang Wan, Christoph Langer, Nuno Bettencourt, Eugenio Martuscelli, Said Ghostine, Ronny R Buechel, Konstantin Nikolaou, Hans Mickley, Lin Yang, Zhaqoi Zhang, Marcus Y Chen, David A Halon, Matthias Rief, Kai Sun, Beatrice Hirt-Moch, Hiroyuki Niinuma, Roy P Marcus, Simone Muraglia, Réda Jakamy, Benjamin J Chow, Philipp A Kaufmann, Jean-Claude Tardif, Cesar Nomura, Klaus F Kofoed, Jean-Pierre Laissy, Armin Arbab-Zadeh, Kakuya Kitagawa, Roger Laham, Masahiro Jinzaki, John Hoe, Frank J Rybicki, Arthur Scholte, Narinder Paul, Swee Y Tan, Kunihiro Yoshioka, Robert Röhle, Georg M Schuetz, Sabine Schueler, Maria H Coenen, Viktoria Wieske, Stephan Achenbach, Matthew J Budoff, Michael Laule, David E Newby, Marc Dewey,

1340 related Products with: Diagnosis of obstructive coronary artery disease using computed tomography angiography in patients with stable chest pain depending on clinical probability and in clinically important subgroups: meta-analysis of individual patient data.



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#31145683   2019/05/15 To Up

A balancing act: PHLPP2 fine tunes AKT activity and MYC stability in prostate cancer.

PTEN loss stimulates prostate tumor progression by sustaining AKT activation. Nowak et al. (2019. https://doi.org/10.1083/jcb.201902048) surprisingly show that the AKT-suppressing phosphatase PHLPP2 promotes disease progression in the context of dual PTEN and p53 loss by increasing MYC stability.
Roxanne Toivanen, Luc Furic

1280 related Products with: A balancing act: PHLPP2 fine tunes AKT activity and MYC stability in prostate cancer.



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#31109340   2019/05/20 To Up

Pre-clinical evaluation of Minnelide as a therapy for acute myeloid leukemia.

There is an urgent need for novel and effective treatment options for acute myeloid leukemia (AML). Triptolide, a diterpenoid tri-epoxide compound isolated from the herb Tripterygium wilfordii and its water-soluble pro-drug-Minnelide have shown promising anti-cancer activity. A recent clinical trial for patients with solid tumors confirmed the safety and efficacy at biologically equivalent doses of 0.2 mg/kg/day and lower.
Bhuwan Giri, Vineet K Gupta, Brianna Yaffe, Shrey Modi, Pooja Roy, Vrishketan Sethi, Shweta P Lavania, Selwyn M Vickers, Vikas Dudeja, Sulagna Banerjee, Justin Watts, Ashok Saluja

2872 related Products with: Pre-clinical evaluation of Minnelide as a therapy for acute myeloid leukemia.

100 assays96 Tests50 assays1,000 tests100Tests100 tests10 assays

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#31046118   // To Up

Intraductal fulvestrant for therapy of ERα-positive ductal carcinoma in situ of the breast: a preclinical study.

Mammographic screening for breast cancer has led to increased detection of ductal carcinoma in situ (DCIS) and a reappraisal of the necessity of aggressive treatment with their attendant toxicities for a preneoplastic lesion. Fulvestrant, a selective estrogen receptor degrader, is very effective in the treatment of estrogen receptor positive (ER+) breast cancer, but delivery by the painful intramuscular (i.m) route is limiting. We hypothesized that intraductal (i.duc) administration of fulvestrant will provide a direct, safe and effective treatment for DCIS. Mice bearing mammary ductal xenografts of ER+, luciferase-tagged MCF-7 breast cancer cells were administered vehicle or fulvestrant i.m or i.duc. I.duc MCF-7-luc tumors in mice treated with fulvestrant i.duc or i.m grew significantly slower than vehicle control. Whole mount analysis and histopathology showed that i.duc fulvestrant achieved significantly larger cancer-free areas. Western blot analysis showed reduced levels of estrogen receptor alpha (ERα) and its downstream targets, c-Myc and Cyclin D1, and increased levels of ERβ, which is known to inhibit ERα function. Immunohistochemical analysis of tumor sections showed that Ki67 and ERα protein levels decreased by 3-fold, and neoangiogenesis was inhibited by i.duc fulvestrant treatment. I.duc fulvestrant also reduced outgrowth of ERα+, autochthonous N-methyl-N-nitrosourea-induced mammary tumors in rats. Overall, we have shown that i.duc fulvestrant was significantly more effective than, or equivalent in action to i.m fulvestrant in two preclinical models of breast cancer. These studies provide evidence for a novel and safe route for fulvestrant therapy of DCIS and prevention of breast cancer. This preclinical study provides a strong basis for conducting clinical trials for DCIS and early breast cancer.
Guannan Wang, Chuang Chen, Priya Pai, Preethi Korangath, Shengrong Sun, Vanessa F Merino, Jingping Yuan, Suping Li, Guangjun Nie, Vered Stearns, Saraswati Sukumar

1828 related Products with: Intraductal fulvestrant for therapy of ERα-positive ductal carcinoma in situ of the breast: a preclinical study.



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