The Chemical Synthesis of Long and Highly Modified RNA using 2'-ACE Chemistry
Poster Oct 01, 2016
Xiaoqin Cheng, Kristina Larson, Letitia Kwok, David Mierzejewski, Shawn Begay, Randy Rauen, Kelly Grimsley, Kaizhang He, Michael Delaney, David Kitchen, Amanda Haas, Melissa Kelley, Anja van Brabant Smith
Due to increased uses of various RNAs in understanding critical structural, functional, and regulatory roles of RNA in biology, rapid, reliable, and cost-efficient methods of RNA oligonucleotide synthesis are in demand. Traditional methods of RNA synthesis based on 2'-silyl (TBDMS or TOM) protection strategies are limited in their ability to construct oligos longer than 40 nucleotides in length (far smaller than important biologically active RNA molecules). A significant improvement in RNA synthesis technology, 5'-silyl-2'-acetoxy ethyl orthoester (2'-ACE) chemistry, results in faster coupling rates, higher yields, greater purity, and superior ease of handling. Using 2'-ACE chemistry, we have developed convenient and efficient protocols to synthesize: (1) long RNA sequences in excess of 100 bases, (2) transfer RNAs (tRNAs) with natural modifications such as pseudouridine, m2A, and m2G, (3) RNA oligonucleotides highly modified with virtually any chemical modification; and (4) long RNA with dual-labeled fluorescent dyes. Our results clearly demonstrate that 2'-ACE chemistry is the method of choice for long RNA synthesis applications.
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
P450 Induction in Cryopreserved Hepatocytes from PXR and CAR Nuclear Receptor Knock-out RatsPoster
The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that regulate the expression of phase I (cytochrome P450s), phase II metabolizing enzymes and transporter genes in response to xenobiotics, including prescription drugs.READ 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.
International Conference on Neurooncology and Neurosurgery
Sep 17 - Sep 18, 2018