School of Science Division of Life Science 24 Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: LI Xingrui / BCB-IRE Course: UROP 1100, Summer Cilia are hair-like cellular organelles that originate from centrioles and extend from the surface of many cells. Despite their diminutive sizes, they are indispensable for signaling, sensory functions, and the overall health of the organism. Defects in ciliary function can lead to a group of diseases called ciliopathies, which can affect multiple organ systems. Primary ciliary dyskinesia (PCD) is one of the most characteristic disorders within this group. To gain a deeper understanding of PCD and to identify genes whose absence or mutation can result in ciliary dysfunction, we employ CRISPR to knock out cilia-related genes reported in other papers. By using super-resolution microscopy, we can observe the changes in the structures and cellular activities of the knock-out cells. Genome Editing by CRISPR Supervisor: LIU Zhen / LIFS Student: WONG Pui Yee / BIOT Course: UROP 1100, Spring UROP 2100, Summer Primary ciliary dyskinesia (PCD) is an incurable genetic disease caused by the mutated gene of mobile cilia. To investigate probable treatment, this project attempts to apply CRISPR system to execute the gene knockout so that the information of inactive or removed genes can be obtained and conducive to understand the mechanism. The feature of altered cilia beating action with edited genes is also observed by diverse microscopy. In current state, the experiments continued and there are another 16 genes, which are also found relating to PCD, targeted and their own gRNAs are also designed in progress and sequenced. In addition, the successfully constructed sample were further purified and analyzed their efficiency. This report aims to state out the present progress and result. CRISPR/Cas9 Analysis of Essential Genes Supervisor: POON Randy Yat Choi / LIFS Student: CHAN Kwing Hei / BIBU Course: UROP 1100, Spring With the aid of a conditional silencing cell line using a combination of CRISPR-Cas9 and degron-tagged cyclin A, AIDCyclin AKO. The study aimed to investigate how varying confluency of the cells affect the growth of AIDCyclin AKO cells. We confirmed that the treatment of doxycycline (Dox) and indole-3-acetic acid (IAA) (i.e. DI) could successfully knockout AID-Cyclin A, which plays an important role in proliferation of the cells. Experiments have shown that more cells were arrest at G2 phase after adding DI in cells seeding at low confluency, suggesting the effect of DI would be strengthened when cells are at low density.
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