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Development of Fluorometric Assay to Detect FTO Binding to Methylated Model RNA
The methylation of the sixth position of adenine (m6A) is a vital epitranscriptomic alteration that has been found to play a vital role in early cell differentiation and embryonic development. The removal of the m6A modification is facilitated by “eraser” protein FTO. The protein FTO (Fat-mass and obesity-related transcript) is a demethylase enzyme whose activity has been recently linked to cancer progression and development. However, little is known about the biophysical nature of its binding to its RNA targets. A TLC-based Mobility Shift Assay with γ32P labeled mRNA which was incubated with FTO resulted in loss of the m6A modification. Additionally, FRET experiments were used to determine the effect of FTO’s cofactors (α-ketoglutarate and Fe (II)) on its binding thermodynamics. FRET experiments also indicated the potential of a phage-display isolated peptide to serve as an inhibitor to FTO’s demethylase activity. Recently, more work has been undertaken to investigate the binding of this peptide to a modified RNA target using Circular Dichroism spectroscopy. - Image
Measuring Organic Field-Effect Transistors
Andrew Shepherd Banner ID: 810962517 Dr. Bjorn Lussem, Physics Department Email : blussem@kent.edu Measuring Organic Field-Effect Transistors Organic field-effect transistors (OFET’s) are of growing interest in the fields of physics and electronics for many reasons. Not only do OFET’s have the ability to be more cost-effective than traditional transistors, they are also very flexible. The flexibility of these transistors creates a wide array of applications for real-world use. OFET’s can be doped with other materials in order to increase the performance of the device. In Dr. Lussem’s lab, doped OFET’s are processed and measured, with electron mobility of specific interest. The transistors are processed by sublimating the materials onto a glass plate, layer by layer. The transistor is moved to the glovebox, where measurements are done to find the electron mobility. Within the pressurized glovebox, a voltage is applied across the transistor. Sensors are connected to a computer program that creates and plots the data points. This data can be analyzed to calculate the electron mobility. The specific compound being measured is DNTT (dinaphtho[2,3-b:2',3'-f]thieno[3,2-b]- thiophene). DNTT is of interest because of other reports being published recently, citing remarkable electron mobility. This high mobility must be corroborated, which is the main reason that this project of processing and measuring DNTT OFET’s was undertaken. - Image
Investigating the role of RsmC C-terminal domain in RNA binding
Ribosomes are the large ribonucleoprotein complexes that are responsible for protein biosynthesis in all organisms. The ribosome’s accuracy regarding its function is heavily controlled by its structural integrity. Ribosomal RNA modifications can influence nearby structures which can then influence the functional efficiency of the ribosome. The focus of the Abey Lab is to understand the ability of nucleotide modifications and their respective modification enzymes in the process of biogenesis of ribosomes. The rRNA nucleotide modification enzyme RsmC is important for the m2G modification at position 1207 in the 16S rRNA. Protein RsmC has an N-terminal domain that functions as the catalytic domain, which is connected to the C-terminal domain through a flexible linker. We hypothesize that the C-terminal domain is important for its binding. We performed site-directed mutagenesis to delete the entire C-terminal domain. We carried out filter binding assays with the mutant and the wildtype protein to determine if C-terminal domain plays a role in binding of RNA.Comparative Analysis of Neutron Star and White Dwarf Star Simulations
Neutron Stars are the remnants of massive stars at the end of their life cycles. They are formed when fusion fuel is depleted, resulting with the inward force of gravity having no resistence. During the collapse, even atoms are unable to maintain their structure, leaving the body of the star as a soup of neutrons. The free eletrons act as a degenerate fermi gas, making them extremely hard to compress, where the resulting outward pressure counters the force of gravity. Once the system is stable, the star contratcs down to a small fraction of its original size with extreme density. In our research, we are building a code to accurately simulate a neutron star and its resulting nuclear interactions at high temperature and high magnetic field. In doing so, we used comparative data to test the this simulation with a corresponding white dwarf code due to their similarities; The compared quantitative aspects were as follows: magnetic field, number density, energy density, pressure, perpendicular pressure, chemical potential, magnetization, and entropy. Overall, it was found that they behaved the same way, and the neutron star simulation is functional.