|Development of an Automated siRNA Screening of Host Macrophages Genes Involved in Mycobacterium Tuberculosis Infection|
Jean-Philippe Carralot, Chang-Bok Lee, Annette Boese, Boris Lenseigne, Auguste Genovesio, Peter Sommer, Thierry Christophe and Priscille Brodin
In order to identify host genes required for M. tuberculosis infection and persistence, we developed a phenotypic cell-based assay in both murine and human cells and adapted it for high throughput and high content screening purposes. Knock-down efficiencies above 80% were achieved in “hard-to-Transfect” macrophage cells. Validation of the assay performed with control siRNAs will be discussed.
|Quantification of siRNA by a Novel Competitive-qPCR Method|
Wei-li, Liu., Mark Stevenson and Len Seymour
We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity
|The Silencing of Multidrug Resistance-Associated Protein 5 by siRNA Complexes|
M.Malinen, E.Mannermaa, T.Ryhänen, E.Kuusela, M.Häkli, M.Yliperttula and A.Urtti
The purpose is to study the role of multidrug resistance-associated protein 5, MRP5 in drug metabolism in human retinal pigment epithelium (RPE). RPE forms the outer part of blood-retinal barrier (BRB) which restricts movements of solutes from systemic bloodstream to the neural retina. The efflux protein, MPR5 is expressed in RPE but its functions are mainly unknown. SiRNA will be tested as a tool to clarify the role of MRP5.
|Two-Dimensional Molecular Profiling of Multiple Myeloma |
Zelena Jana, Konecna Hana, Zdrahal Zbynek, Penka Miroslav and Hajek Roman
We have compared two different solubilization buffers, we also evaluated protein precipitation with ethanol and optimized 2-DE conditions for human myeloma proteins.
|2-DE Analysis of Breast Cancer Cell Lines MDA-1833 and MDA-4175 with Distinct Metastatic Organ-Specific Potentials and Comparison with Parental Cell Line MDA-MB-231|
Irena Selicharová, Miloslav Šanda, Lucia Svitekova, Sujata Suraswat Ohri, Aruna Vashishta, Martin Fusek and Václav Vetvicka
Breast cancer has diverse metastatic behavior. Subpopulations of cells with enhanced metastatic abilities either to bone (1833) or to lung (4175) have been isolated by the group of J. Massague by in vivo selection of MDA-MB-231 cells. We performed the 2-DE analysis of the cell lines. We have found and identified 11 differentially expressed protein spots.
|Expression Profiling of CD34+ of Peripheral Blood of Patients with Lymphoma During ex vivo Granulocytic Differentiation|
Irena Koutna, Lenka Tesarova, Martin Klabusay, Petr Krontorad, Michal Strehovsky, Martina Peterkova and Viera Hrabcakova
On the basis of global expression data analysis we have concluded that population of CD34+ cells from patients with lymphoma shows different expression status during ex-vivo differentiation. And thus we assume that CD34+ population is not suitable for autologous transplantation. We have used cDNA microarrays technology to obtained precise expression profiles.
|MS-Xelerator™: Advanced Algorithms for LC/MS Data Processing Applied to Biomarker Discovery, Differential Analysis and Quantitative Proteomics|
LC-MS based proteomic experiments are used to compare complex biological samples across multiple conditions. Fast, powerful computational tools are needed to explore and detect differences in the areas of Expression Proteomics and Biomarker Discovery. In general, specialized steps are necessary to solve these difficult problems (binning, alignment & normalization, peak picking, relative quantitation, etc.). MS-Xelerator is a collection of software tools dedicated to all of the above tasks.
|Caveolin-1 Expression as a Possible Biomarker in Pancreatic Cancer Diagnosis|
C. Tanase, E. Raducan, L. Albulescu, E. Codorean, M.I. Nicolescu, D.I. Popescu, M.L. Cruceru and A.C. Popa
Caveolin1 (Cav-1) function either as a tumor supressor or as a promoter of metastasis. Overexpresion of cav-1 was correlated with: tumoral grading, proliferration markers (Ki67, p53), serum tumor markers (CEA, CA19.9) and angiogenic markers (VEGF, bFGF).
|Antigen-Specific Delivery of siRNA Against Eucaryotic Elongation Factor 2 by Rationally Designed Bivalent Aptamer-siRNA Transcripts |
nga Neef, Ulrich Wüllner, Andreas Eller, Michael Kleines, Rainer Fischer, Mehmet Kemal Tur and Stefan Barth
Here we show specific cytotoxicity resulting from siRNA-induced silencing of EEF2, as well as specific delivery to PSMA-expressing prostate cancer cells. Increasing the valency of the aptamer resulted in enhanced cytotoxicity compared with the monovalent constructs. The results presented here demonstrate the usefulness of multivalent aptamer-based delivery vehicles for siRNA therapeutics.