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RNAi
Phenotypic loss-of-function RNAi screens with complex lentiviral-based shRNA expression libraries that target and silence several thousand genes provide a realistic and workable approach to identify genes that functionally modulate a cellular response such as viability of cancer cells or apoptosis. As long as the shRNA libraries are properly constructed so that hairpin representation is well characterized and reasonably constrained, and changes in shRNA representation in selected vs.
High-Throughput Screening
A suitable High-Throughput and High-Content Screening strategy has a major impact on drug discovery. The use of the most advanced technologies is a major prerequisite for successful screening.
Gene Targeting
A major challenge facing the development of PI3K-targeted pharmaceutical agents is the lack of patient-relevant, in vitro model systems. Studies into the function of oncogenic mutations are commonly carried out in cell line panels that are mutant or wild-type for the gene of interest, however interpretation of these studies can be confounded by additional genetic differences between each cell line background, calling into question the reliability of the results.
Epigenomics
Histone deacetylases (HDACs) are a class of enzymes that remove acetyl groups from ε-N-acetyl lysine amino acids on histones and other proteins. The removal of acetyl groups serves to increase the positive charge of histone tails, encouraging binding between histones and the DNA backbone, and preventing transcription.
Epigenetics
Post-translational modifications of histones are among the epigenetic mechanisms that can affect chromatin structure and function. Disruption of epigenetic processes can lead to altered gene expression and malignant cellular transformation.
Epigenetics
Several assay methods have been developed for quantifying the activity of histone deacetylases (HDACs and sirtuins), histone methyltransferases (HMTs), and histone demethylases (HDMs). These include radioactive assays, enzymelinked immunoassays (ELISA), mass spectrometry, and enzyme-coupled detection of fluorescent peptides or reaction co-products (e.g.
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