School of Science Division of Life Science 25 CRISPR/Cas9 Analysis of Essential Genes Supervisor: POON Randy Yat Choi / LIFS Student: ZHU Jingjun / BIBU Course: UROP 2100, Fall The presence of centrosome amplification is a hallmark of cancer cells. Cancer cells with supernumerary centrosomes adopt a mechanism called centrosome clustering to maintain the ability of constant proliferation. Some cancer cells with extra centrosomes can achieve pseudo-bipolar mitotic division through centrosome clustering and produce viable daughter cells. Many studies have shown that kinesins are essential in regulating centrosome clustering in cancer cells. By studying the specific functions of different kinesins in the regulation of centrosome clustering, it is possible to develop new directions for cancer treatment. My previous studies validated the knockout efficiency of CRISPR plasmids targeting KIF4A, KIF10, and KIF23 genes. This semester, the objective of my experiments was to further validate the functions of these three kinesins in regulating centrosome clustering in HeLa cells. Live-cell imaging analysis revealed that depletion of these kinesins may affect centrosome clustering. Further studies can focus on reproducing the same experiments to confirm the functions of these kinesins. Development of an Algorithm to Analyse Live-cell Imaging Data Supervisor: POON Randy Yat Choi / LIFS Student: PAN Yunqing / DASC Course: UROP 1100, Spring UROP 1000, Summer This report outlines improvements and new developments in a cell mitosis analysis program. Enhancements include a refined preview feature that iteratively searches for a usable cell ID to ensure reliable previews. The user interface (UI) has been streamlined to resolve structural issues and improve clarity. The new "percell version" comprises two Python files for UI and data processing, facilitating a three-step workflow: data reading, cell object identification, and result selection. Users can manually select mitosis results based on STD curves, with error checks for file format issues, peak detection, and frame numbers. These updates should be able to enhance the program’s accuracy and user experience.
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