School of Science Division of Life Science 25 Targeting Mitotic Regulators in Cancer Cells for Potential Treatment Supervisor: POON Randy Yat Choi / LIFS Student: THEN Claudia Regina / BCB Course: UROP1100, Spring Chinese Hamster Ovary (CHO) cell is frequently used for the production of recombinant protein. There are several steps to creating a culture of CHO cells expressing recombinant protein. First, the gene of interest is amplified using polymerase chain reaction (PCR). Then, the amplified DNA fragment and recipient plasmid DNA are digested using restriction enzymes to create complementary ends. After that, the digested DNA fragments are isolated using gel purification. The gene of interest and plasmid DNA are then connected using ligation procedure to make a complete plasmid. Lastly, the plasmid is transfected into CHO cell culture. This report will describe in more detail the standardized version for each procedure as the experiments have not yet been performed in actuality. CRISPR/Cas9 Analysis of Essential Genes Supervisor: POON Randy Yat Choi / LIFS Student: BYUN Seung Min / BIBU Course: UROP1100, Fall DHX9 is an ATP-dependent RNA helicase that may function in the cell cycle. I conducted a project on transfecting DHX9 CRISPR into a HeLa cell to confirm its effect of knockout as well as its potential function in the cell cycle. Using calcium phosphate-mediated transfection and Western blotting, I showed that the CRISPR was not effective in knocking out the gene of interest. Additional constructs of DHX9 CRISPR were designed for future experiments. CRISPR/Cas9 Analysis of Essential Genes Supervisor: POON Randy Yat Choi / LIFS Student: KIM Jimin / BIOT Course: UROP1100, Spring UROP2100, Summer Cells can be synchronized at specific phases through different ways such as chemical blockade, serum deprivation, centrifugal elutriation, etc. Among, RPE1 cells are increasingly widely being used as models in biomedical research. However, compared to HeLa, which is the most commonly used human cell line, an effective way to synchronize RPE1 cells has not been studied yet. Therefore, this report aims to examine a way to achieve cell synchronization of RPE1 cells by attempting different methods, including serum starvation and the utilization of Lovastatin. After conducting FACS with the samples, at which phase of the cell cycle the cells were trapped was explored.
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