Title: Role of SATB2 in serrated colorectal cancer development
In this study, we will investigate the role of a candidate tumour suppressor gene in colorectal cancer development. This will involve molecular work on in vitro and tumour tissues harvested from mice experiments. Techniques will include molecular cloning, PCR/qPCR, immunoblot and flow cytometry.
Title: Investigating molecular players of innate immune response
The innate immune response is the first line of the defense system of our cells against pathogens. Upon encountering a foreign antigen, our cells initiate a series of complex signaling cascades to effectively express the required tools to clear the foreign invaders. We are studying sequential molecular events to understand how this evolutionarily conserved system works in a timely manner. We identified telomerase protein, an enzyme that is known to maintain telomeres, as one of the regulators of innate immune response and aimed to explore the molecular details and its involvement in host-pathogen interactions.
Title: Bioengineering of synthetic internal ribosomal entry site (IRES) for efficient bicistronic expression for application in genome editing and gene therapy
In contrast to polycistronic transcripts in prokaryotes, eukaryotic mRNA generally only express a single gene. Several strategies, such as ribosomal skipping peptide sequences or internal ribosomal entry sites (IRES), have been used to create bicistronic transcripts. However, their efficiency has been underwhelming. Inspired by naturally existing sequences from viruses and human genes, we will screen and determine small nucleic acid sequences for efficient use in viral production, genome editing constructs, and gene therapy.
The projected difficulty is suitable for undergraduates while providing a sufficient challenge that facilitates the student’s learning and growth. The student will be exposed to a research laboratory environment and learn vital molecular biology methods (PCR, cell culture, molecular cloning, bacterial culture, DNA and RNA preps, western blot, flow cytometry, transfection, viral transduction, etc.). The experience and mentorship from this project will prepare the student for a successful FYP and eventual employment or graduate degree pursuit.
We are looking for students who:
1) is highly motivated and passionate about learning;
2) takes ownership of their work and progress;
3) is adaptable in a fast pace environment; and
4) being a contributive team member.
Objective:
1) Molecular cloning of human 5’ UTR and viral sequences in plasmid constructs
2) Screen these sequences for IRES activity and the effect on upstream and downstream genes
3) Create a truncation construct to screen for a minimal functional portion of the IRES
4) Application of these characterized IRES in lentiviral and AAVs constructs for efficient reporting of gene expression and transduction
Field of Research:
Molecular Biology, Bioengineering, Biochemistry, Cell biology, Synthetic biology, Genome editing
Methods/Techniques:
- Polymerase chain reaction (PCR)
- Molecular cloning via gene fragments assembly
- Bacterial transformation
- Bacteria culture
- DNA, RNA, Protein preps
- Transfection
- Transduction
- Cell Culture
- Flow cytometry
- Western blot
- Quantitative PCR
Relevance:
Successful engineering of short and efficient IRES sequences would serve as excellent tools to report gene delivery. Furthermore, multicistronic transcript would help alleviate the size limitation of common gene therapy vectors such as AAVS.
Title: Exploiting ferroptosis to eliminate colorectal cancer cells with KRAS mutations
Ferroptosis is an iron-dependent form of regulated necrotic cell death triggered by oxidative degradation of lipids in membrane bilayers and subsequent irreparable damages to the plasma membrane. This fatal chain of reactions occurs when the downregulation of key metabolic determinants creates an imbalance between the production of free radicals and antioxidants. Notably, lipid peroxidation is observed following:
i) the depletion of intracellular glutathione (GSH) via the inhibition of cell membrane transporters such as the cystine/glutamate transporter;
ii) the blockade of intracellular antioxidant enzymes (such as glutathione peroxidase GPX4);
iii) increased labile iron pools.
Interestingly, whereas the contribution of ferroptosis in development and physiological homeostasis remain largely obscure, induction of ferroptosis has been convincingly established as a potentially important strategy to improve cancer treatment. In particular, colorectal cancer (CRC) cells harbouring oncogenic KRAS mutations are exquisitely sensitive to ferroptotic activators such erastin and RAS-selective lethal 3 (RSL3). CRC is the third most deadly cancer and the second leading cause of global cancer mortality worldwide. Despite the development of diagnostic and therapeutic strategies, most CRC patients are diagnosed at an advanced stage and CRC remains one of the most severe human cancers due to resistance to treatment. Hence, this project will aim at understanding the basic mechanism underpinning CRC cells sensitivity to ferroptosis through KRAS-mediated metabolic reprogramming in order to establish novel therapeutic opportunities to treat incurable CRC.
Title: Characterizing hepatocellular carcinoma at spatial resolution
In this project, we will utilise clinical samples to perform cutting edge single cell and spatial transcriptomic to reconstruct the ecosystem that make up hepatocellular carcinoma.
Title: Identification of novel targetable vulnerability in IDH wild-type glioblastoma
We have identified LOC a novel RNA as an important component of NFκB signalling. Next, we aim to
a) understand the mechanism by which LOC works to regulate NFκB
b) identify the context in which it works and
c) delineate if blocking LOC can be developed as a therapy in future.
This knowledge will provide therapeutic options to overcome drug resistance in GBM and other cancers.
Title: Study an interesting gene in the development and progression of colorectal cancer via regulating ferroptosis
Colorectal cancer (CRC) is the third most prevalent cancer and the second leading cause of cancer mortality. Although patients have more treatment options for CRC patients, the prognosis is poor for metastatic CRC patients. Ferroptosis, a newly discovered form of regulated cell death driven by iron-dependent excessive lipid peroxidation, has been implicated in the development and therapeutic responses of various types of tumors. In our project, we mainly focus on elucidating the mechanism of ferroptosis in the development and progression of CRC to explore potential therapeutic targets for CRC patients.
Title: Genome wide CRISPR screening of metastatic tumors
We will set up CRISPR screen to identify the candidates involved in cancer metastasis and further, validate and investigate mechanistic role of these candidates in tumor progression.
Title: Small molecule screening against regualtor of hTERT promoter
In this project, we will make use of reporter cell lines to screen small molecule libraries against the regulators of hTERT promoter and then validate their efficacy on cancer progression invitro.
