Automation of a Generic Fluorescence Methyltransferase Activity Assay
Poster Feb 16, 2012
X. Amouretti, P. Brescia, P. Banks, G. Prescott, Meera Kumar
Methylation is known to be a ubiquitous covalent modification involved in regulation of a diverse range of biomolecules. Histone methyltransferases (HMTs) are of particular interest as drug targets as histone methylation is linked to certain disease states, including a wide variety of cancer types. A high-throughput screening (HTS)-ready, universal methyltransferase activity assay was recently developed based on competitive fluorescent polarization immunodetection of AMP, formed from the methyltransferase (MT) reaction product S-adenosylhomocysteine in a dual enzyme coupling step. This work demonstrates automation of the assay in a 384-well format suitable for HTS.
Characterization of a Type 2 diabetes-associated islet-specific enhancer cluster in STARD10 by genome editing of EndoC-βH1 cellsPoster
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE
Psychiatric Risk Gene Cacna1c and Early Life Stress: Potential Gene-Environment interactions?Poster
Early life stress (ELS) is highly associated with development of psychopathology
and mood disorders in adulthood. Genetic studies have identified variation in the gene calcium voltage-gated channel subunit alpha1C (CACNA1C) to increase risk for several psychiatric disorders. This poster assessed the expression of Cacna1c following prepubertal stress.
Knockout of microRNAs Using the CRISPR-Cas9 System with Paired Synthetic crRNAsPoster
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE