School of Science Division of Life Science 29 Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: WONG Pui Yee / BIOT Course: UROP 3200, Spring UROP 4100, Summer Retinitis pigmentosa GTPase regulator (RPGR) is a critical protein associated with cilia, characterized as hairlike structures found in various organs throughout the body. Mutations in RPGR genes contribute to 70-90% of retinal pigmentosa (RP) cases, a hereditary eye disorder that causes blindness. In addition, some patients with RPGR mutations exhibit predisposition to primary ciliary dyskinesia (PCD), a rare airway disease. However, their relationship and pathway are still not fully understood. Our previous work showed the abnormal F-actin accumulation in RPGR knockout (KO) cells, but it is unclear whether actin dynamic changes is the cause of the RPGR-RP and RPGR-PCD. This project aims to further investigate the role of actin dynamics in the diseases caused by RPGR defects. Therefore, we can reveal the disease mechanism correlated with RPGR mutations and identify downstream targets for therapeutic interventions. Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: YUEN Tsz Man / BIOT-AB Course: UROP 1000, Summer TRPM8 is best known as a cold receptor. Since external temperature can impact respiratory health, it’s suggested that there’s a link between the protein and certain respiratory diseases, especially those involving defective cilia in the respiratory tract. To test the hypothesis, the Trpm8 gene is knocked out in human ciliated cells using CRISPR, and their ciliary morphology is compared with that of control samples. This paper mainly describes the methodology used and the principles behind it. Since an unexpected delay has occurred, this study has just reached the “lentivirus packaging” stage. Nonetheless, significant progress has been made as plasmids encoding different gRNAs have all been successfully constructed and acquired. Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: ZENG Qianhui / BCB Course: UROP 1100, Fall This report chronicles my journey and progress in a laboratory dedicated to gene editing using CRISPR. Beginning under the help of the senior, I was introduced to the first view of laboratory research, getting a closer look at advanced biochemistry lab techniques, including DNA annealing, enzyme digestion an plasmid transfection. Over time, I have grown up from a newcomer to an independent assistant, arranging my own experiment schedule, actively contributing to the lab’s projects and developing my own research plan. This experience has not only deepened my technical expertise but also fostered my ability to work in a fast-paced research environment. The report outlines the methods employed, results achieved, and the progress I have made.
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