|Gene Expression from Pseudourine and 5-Methylcytidine Modified Messenger RNA|
Jiehua Zhou, Maggie L. Bobbin, Julie R. Escamilla-Powers, Anton P. McCaffrey and John J. Rossi
Study objective was to develop methodologies for gram scale synthesis of messenger RNA (mRNA) for gene therapy applications, as well as scalable purification methods that yield highly expressed, persistent and non-toxic mRNA.
|Random Homozygous Gene Perturbation (RHGP) as a Tool for Target Discovery and Validation|
Wu-Bo Li and Michael Goldblatt
Random homozygous gene perturbation (RHGP) can identify and validate any host (cellular) gene target that directly causes a desired phenotype without requiring prior knowledge of the target. The central feature of RHGP is a unique lentiviral-based genetic element, known as a gene search vector (GSV) designed to interrogate the entire genome and identify target genes that cause the phenotype of interest.
|Simultaneous RT-qPCR Measurement of 1718 Long Non-Coding RNAs|
Pieter Mestdagh, Barbara D’haene, Jan Hellemans and Jo Vandesompele
Massively parallel RNA-sequencing revealed that the human genome is pervasively transcribed, resulting in the production of thousands of non-coding RNA transcripts.
|Advanced Copy Number Variant Analysis with qbasePLUS 2|
Barbara D’haene, Jo Vandesompele and Jan Hellemans
Copy number changes under the form of deletions and duplications are known to be involved in numerous human genetic disorders. Moreover, each individual’s genome embodies several copy number polymorphisms of various sizes which are thought to contribute to normal phenotypic variation and susceptibility to multifunctional disease.
|ChIP-qPCR and qbasePLUS Jointly Identify a MYCN Activated miRNA Cluster in Cancer|
Barbara D’haene, Pieter Mestdagh, Daniel Muth, Frank Westerman, Frank Speleman and Jo Vandesompele
This study applies ChIP-qPCR tp assess binding of transcription factor MYCN to miRNA cluster 17-92, to positive control target MDM2, and to a negative control target region.
|Development of an Automated Platform for HT Cloning and Expression|
Stefano Bonacci; Sara Iozzi; Scilla Buccato; Manuele Martinelli; John Telford; Domenico Maione; Roberto Petracca
Biomolecular protocols covering the whole cloning process were implemented in liquid handler robots. When compared to the manual approach, it was found that automation significantly speeds up HT cloning.
|Volume-Related Inhibitors Standardization for Reverse Transcription Quantitative Polymerase Chain Reaction Experiments|
Pascal Pugniere, Sebastien Banzet, Thomas Chaillou, Catherine Mouret and Endre Peinnequin
This poster addresses the reliability of qPCR data and its dependence on technical variations. The proposal is that constant volume of RNA extract can improve reliability of RT-qPCR.
|Defining off-target cleavage in a pair of Zinc Finger Nucleases|
K. Mukherjee, D. Carroll
This study looks at off-target cleavage of Zinc Finger Nucleases (ZNFs) in Drosophila in an attempt to analyze potential cleavage spots, with a view to designing more efficient ZFNs.
| Gene Expression Profiling: qPCR Toolkit for Quality Control|
Švec D., Jacobsson J., Sjöback R., Kubista M.
TATAA Biocenter explain how they developed and optimized high-throughput gene expression qPCR with ValidPrime quality control, compensating for inter-run variations.