School of Science Division of Life Science 33 G Proteins and Their Regulators in Cancer Biology Supervisor: WONG Yung Hou / LIFS Student: ZHANG Huixian / LIFS Course: UROP 1100, Fall Among the guanine nucleotide-binding proteins (G proteins), Gαi1 protein function as an inhibitor of adenylyl cyclase (AC) transferring ATP to cyclic adenosine monophosphate (cAMP) and pyrophosphate. In this study, we aimed to identify the key sites responsible for Gαi1 protein functioning by mutating four specific residue groups to different amino acids, and transferring them into mammalian cells. To isolate the effect of mutated Gαi1 protein, we inhibited other endogenous Gαi1 protein functioning using pertussis toxin (PTX). For reflecting the protein’s inhibitory effect, we utilized a fluorescent assay to measure intracellular cAMP level whose decrease can indirectly indicate the inhibitory effect. Our findings provide insights into the functioning mechanism of Gαi1 protein, especially how much the targeted residues contribute to the Gαi1-AC reaction, which may have value for further drug development and other application. Employ Mathematic Modelling to Study Gene Networks Regulating Stem Cell Development and Eye Diseases Supervisor: XIE Ting / LIFS Co-supervisor: YANG Can / MATH Student: GAO Shuyang / MATH-STAT Course: UROP 1100, Spring In this progress report, we study Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) to investigate the specific sites of binding events of three serine/argininerich splicing factors (SRSFs): SRSF2, SRSF3, and SRSF10. Our data pre-processing workflow uses FastQC and Cutadapt, alignment with Bowtie, and cluster generation with PARalyzer. The binding sites were annotated using two genome references. Our results revealed significant overlaps in the binding sites of SRSF3 and SRSF10, with a smaller overlap observed with SRSF2 by using a Venn diagram. Specifically, the shared common overlap among all three factors comprised 13.95% of SRSF2, 10.84% of SRSF3, and 24.17% of SRSF10. The report also offers three strategies through bar charts, pie charts, and box plots on different aspects of the binding sites for further study. Employ Mathematic Modelling to Study Gene Networks Regulating Stem Cell Development and Eye Diseases Supervisor: XIE Ting / LIFS Co-supervisor: YANG Can / MATH Student: LAM Hoi Kei / COSC Course: UROP 1100, Spring RNA editing is the alternation of RNA sequences through insertion, deletion or substitution of nucleotides (Brennicke, Marchfelder, Binder, 1999). The most common type to RNA editing in mammals is Adenosineto-Inosine (A-to-I). Recently, it is found that the number of editing sites and editing level of DROSHA knockdown cells decreases in some particular cells (Quinones-Valdez et al., 2019). However, the effect of DROSHA and DICER on other human cells are still unknown. In this project, we analyze RNA sequences in human cells upon DROSHA knockdown, DICER knockdown and normal condition to investigate their effect to RNA editing and number of editing sites.
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