Specificity and Functionality of microRNA Inhibitors
Poster Mar 10, 2015
Barbara Robertson, Andrew Dalby, Jon Karpilow, Anastasia Khvorova, Devin Leake and Annaleen Vermeulen
Highly potent microRNA (miRNA) inhibitors are valuable tools for elucidating the roles of miRNAs and their targets. Although it is known that pairings between endogenous miRNAs and their natural targets in animals generally involve base pair mismatches, studies of how mismatches between endogenous miRNAs and artificial inhibitor targets might affect inhibitor specificity and functionality have been very limited. Using a Luciferase reporter system we have investigated the specicity of miRNA inhibitors. We first confirmed significant levels of cross-reactivity among the closely related let-7 family members. Subsequently,
a systematic study of mismatches incorporated into inhibitors of single-family-member miRNAs identified two regions that are important for overall inhibitor functionality. Our findings indicate that features important for natural miRNA target recognition also appear to be important for inhibitor specificity. Understanding the specificity of inhibitors allows for better interpretation of inhibitor activity in endogenous systems.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE
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