|Heterochromatin structure is induced by siRNA targeting HIV-1 promoter region |
K Suzuki1, H Lim1,3, T Ishida2, T Watanabe2, D Cooper1,3, A Kelleher1,3
We transfected siRNA targeting HIV-1 promoter region into a cell line productively infected with HIV-1. ChIP analysis revealed that the RNA duplex induced transcriptional gene silencing and enrichment of Ago1, H3K9me2, and HDAC1 in HIV promoter region to form heterochromatin structure. The data indicates RNA duplex induces the latent phase of HIV infection, since the chromosome formation is very similar to the state of HIV latently infected cell lines
|Efficient downregulation of the lung liquid clearing gammaENaC subunit by RNAi|
Nihal Yueksekdag, Marei Drechsel, Christa Schmidt, and Josef Rosenecker
CF is caused by mutations in the gene encoding for CFTR. CFTR functions as chloride channel on the apical membrane of epithelia thereby regulating the transport of chloride and also sodium ions indirectly. It seems that the regulation of ENaC fails due to the mutated CFTR protein. And it is assumed that ENaC plays a role in the pathogenesis of chronic lung disease in CF-patients.
|In Vitro Potency of Chemically Modified siRNAs Against TNF-alpha|
Sandra M. Ocampo, Anna Aviño, Santiago Grijalvo, José C. Perales and Ramón Eritja
The execution of succesful RNAi experiments depends upon multiple factors: 1. Design and identification of effective and specific siRNA sites 2. Enhancement of pharmacocinetic properties 3. Delivery of siRNA. Therefore we have designed, prepared and functionality tested a variety of chemical modifications. These have been tested in HeLa cells to ascertain wether they influence RNAi inducing activity. We have chosen tumor necrosis factor (TNF-alpha) as a target.
|Critical factors for successful RNAi experiments in primary cells and hard to transfect cell lines|
Markus Zumbansen1, Nicole Spottke1, Sheila Offizier1, Allison St. Amand2, Devin Leake2, Ludger Altrogge1, Meike Weigel1, Sandra Domzalski1, Dietmar Lenz, and Herbert Müller-Hartmann1
The amaxa Nucleofector® Technology is a well established method for effective, non-viral transfection of any nucleic acid substrate into hard-to-transfect cells, especially suspension and primary cells. With the
Nucleofector® 96-well Shuttle® system high throughput applications such as siRNA-library screenings have become amenable for the first time in these cell types. This renders target validation and identification possible
in cell types highly relevant for biomedical research. Here we dis
|Highly Efficient High-Throughput Transfection|
Markus Zumbansen1, Allison St. Amand2, Devin Leake2, Ludger Altrogge1, and Herbert Müller-Hartmann1
Successful RNAi experiments and large-scale siRNA screens require efficient delivery of highly functional and specific nucleic acids including siRNA oligonucleotides, shRNA vectors, or micro RNAs into an appropriate cell system. Cell types relevant for immunological research, such as primary T cells and several suspension cell lines, are poorly accessible using reagent-based transfection approaches.
|Automated High Throughput Nucleofection®|
Claudia Merz, Andreas Schroers, Eric Willimann
Using primary cells for RNAi based applications such as target identification or – validation, requires a highly efficient transfection technology in combination with a reliable and robust automation system. To accomplish these requirements we integrated the amaxa 96-well Shuttle® in a Tecan Freedom EVO® cell transfection workstation which is based on Tecan’s Freedom EVO® liquid handling platform and include all the necessary components and features for unattended cell transfection.
|DOCKING STUDIES OF A NEW HETEROCYCLIC METHYLTHIOMORPHOLIN PHENOLS DERIVATIVES AS ANTIHYPERTENSIVE DRUGS WITH ACE TARGET.CASE FESCDIPINE II |
1V. Vázquez,1O. Vázquez, 1,2M.A.Balboa, 1A. Ma. Velázquez, 1V.H. Abrego, 1B.Camacho, 1L. Martínez, 1L.A. Torres, 1A. Ramírez, 1I.Martínez, 3R.López-Castañares, 1S.Díaz-Barriga, 4J. Bocanegra, 4Froylan Gómez-Laguna, 1F. Olvera, 1E.Angeles.
As part of the Drug Design in Medicinal Chemistry Program of the UNAM, new methylthiomorpholinphenol compounds with cardiovascular effect, were studied. One of them, FESCDIPINE II, was an excellent antihypertensive drug, that has low toxicity and preliminary studies indicate that the Angiotensin-converting enzyme (ACE) system is the biological target of this compound.
|Use of gamma scinitigraphy to understand inhaled device/formulation variables on delivery efficiency and|
Peter Scholes and Karen Jones
Systemic delivery of both small molecules and macromolecules via inhaled therapies is an area of significant ongoing research1. The pulmonary route offers the physiological benefits of a highly vascularised, large surface area for absorption which can promote high bioavailability and a rapid onset of action. For biomolecules such as peptides, proteins and nucleic acid derivatives, inhaled drug delivery can also provide a viable alterative to intravenous administration.
|Quantifying the Impact of a Drug on Gastric Emptying: Measuring the Pharmacodynamic Effect in Clinical Trials|
A Rankin, M Paterson, A Connor
Many drug classes are known to alter the rate of gastric emptying. Whilst there is no specific regulatory guidance requiring the impact of drugs on GE to be measured, it is important to fully understand the mode of action and the relationship between the pharmacokinetic profile and the pharmacodynamic response.