UROP Proceedings 2020-21

School of Science Division of Life Science 30 The Rules of Packaging Fat in Cells Supervisor: MAK Ho Yi / LIFS Student: MA Chengkun / BCB Course: UROP2100, Spring tmem-120 is a gene identified in a genetic screen for lipid droplet expansion suppressors. It was shown that tmem-120 loss-of-function mutation could affect fatty acid accumulation to suppress lipid droplet expansion. The first part of this UROP project aims to produce transgenic worms that carry GFP reporters responsive to TMEM-120 activity, which is a continuation of the previous UROP project. The second part of this project intends to test a newly improved technique, the auxin-inducible degron 2 system, in C. elegans. Ideally, this technique allows efficient and reversible protein degradation in response to the plant hormone auxin. An approach similar to the first part is adopted to generate transgenic animals for testing. Characterization of Novel Cell Cycle Regulators in Cancer Cells Supervisor: POON Randy Yat Choi / LIFS Student: CHEUNG Lo Sha / BICH Course: UROP1100, Fall As a member of MYC family, MYCN gene amplification has been shown in multiple human tumors, including a severe form of neuroblastoma. In this research project, by deploying CRISPR-Cas9 technology, different sequences are targeted, causing specific sites of double-strand break (DSB) on MYCN gene, inducing deactivation of MYCN genes. Subsequently, cell cycle stages were examined. A CRISPR targets exon 3’s open reading frame of MYCN was found to be able to affect cell cycle in HtTA1 N-Myc #15 cells. Since many cancers are related to abnormal cell cycle, the study of MYCN related pathway can provide insights for therapeutic development targeting MYCN gene. CRISPR/Cas9 Analysis of Essential Genes Supervisor: POON Randy Yat Choi / LIFS Student: RAG Adwika / SSCI Course: UROP1000, Summer Knockout of genes can be led to a more molecular based and specific treatment for cancer patients. One gene suggested is SPC24. Using auxin-inducible degron (AID) system to conditionally degrade necessary proteins, for which doxycycline and IAA are added to these cells to observe if they undergo mitotic arrest and cell death. Mainly done through immunoblotting and mitotic index calculations, it was observed that some degree of apoptosis was observed but it was not as comparable as other prevalent chemotherapy drugs such as Taxol or Nocodazole (Noc). Furthermore, there was an attempt to see the best dosage of these chemotherapy drugs due to their high toxic side effects, but after serial dilutions, the mitotic arrest and cell death observed through immunoblotting showed that these dilutions are not enough to cause complete death and have reduced efficacy.