|A Novel Multiplexed Digital Gene Expression Technology|
Gary K. Geiss1,#, Roger Bumgarner2, Brian Birditt1, Timothy Dahl1, Naeem Dowidar1, Dwayne L. Dunaway1, H. Perry Fell1, Sean Ferree1, Renee D. George1, Tammy Grogan1, Jeffrey J. James1, Malini Maysuria1, Jeffrey D. Mitton1, Paola Oliveri4, Jennifer L. Osborn3, Tao Peng2, Amber L. Ratcliffe1, Philippa J. Webster1, Eric H. Davidson4, and Leroy Hood5
We describe a novel platform, the nCounter Analysis System, for sensitive, highly multiplexed, digital gene expression analysis based on NanoString’s novel molecular barcoding technology. Detection of individual mRNA molecules using an assigned sequence of six different fluorescent spots per probe are detected, and then the number of times that code sequence appears in a sample are counted.
|Oligonucleotides with LNA and targeting of biologically important RNAs|
Peter Guterstam and Ülo Langel
Activity of RNA-targeting antisense oligonucleotides increases when introducing LNA monomers, implying that an 18 nucleotides long LNA/2OMe mixmer splice-switching oligonucleotide can be shortened to 12mer and have similar activity and specificity as an 18mer 2OMe oligonucleoyide. Positioning of LNA monomers has to be carefully considered when utilizing the potent LNA monomers in RNA-targeting antisense oligonucleotides.
|A comparison of siRNA activity predictors using advanced regression techniques|
Simone Sciabola, Qing Cao, Theresa Johnson, Lingling Shen, Robert Stanton, Xiaoyu Jiang, Simon Xi, Jason Hughes, Daniel Caffrey, Shobha Potluri, Steven Haney, Johnathan Cyr, Jeremy Little
Although considerable progress has been made recently in understanding how gene silencing is mediated by the RNAi pathway, the design of effective sequences is still a challenging task. To tackle the efficacy challenge and further improve accuracy for prediction of siRNA potency, we performed a comparison of advanced regression techniques and new in-house generated descriptors in the generation of siRNA efficiency models.
|miRNAs in Treating Cardiomyopathy |
The study aims to design antigomirs against miRNAs involved in Cardiomyopathy. Potential miRNAs involved in the down regulation of certain important genes during this disorder have been identified. All reported miRNAs were scanned using an algorithm against these genes. At three step protocol was followed to take care of false positives and false negatives. Further, HL-1 cells (cardiomyocytes) are been transfected by anti-miRNAs for confirmation.
|qPCRas a primary screen in drug discovery|
GauravJaggi, Frank Boeckler, Andreas Joerger and Alan Fersht
We report the use of qPCR technique to follow the thermal unfolding of proteins by the binding of the dye SYPRO Orange, and exploit its potential as a robust and high-throughput primary screen for small molecule drug discovery.
|Targeting Inflammatory Cytokines Using Adenoviruses: gene delivery of biological therapies in ovarian cancer|
Michael A. Salako, Hagen Kulbe, Iain A. McNeish, Frances R. Balkwill
Constitutive TNF-alpha expression is characteristic of the malignant ovarian surface epithelium. Adenoviral mutants hold great promise as gene therapy vectors but their efficacy is hindered by an inflammatory cascade orchestrated by TNF-alpha. We found that delivering TNF-alpha shRNA to ovarian cancer cells using oncolytic adenoviruses could reduce the inflammatory cascade generated by adenoviruses and also had direct anti-tumour activity on the cancer cells.
|RNA interference in Manduca sexta induced by double-stranded RNA feeding|
Isabel Gomez, Alejandra Bravo and Mario Soberon
We used a double-stranded RNA feeding approach to silence cadherin-like gene: a protein reported as receptor in several lepidopteran insects for the Cry toxins synthesized by Bacillus thuringiensis. We analized the protein expression from dsRNA-fed and untreated laravae on western blott. M. sexta larvae that ingested dsRNA directed against cadherin exhibited a dramatic suppression of the protein and are resistant to the effect of the Cry1Ab toxin.
|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.