PhD studentship (Dr. Kumaran Narayanan) (1)
Project Title: Genome Engineering in Mammalian Cells Using Phage N15 Protelomerase
Description of the project:
Phage N15 has the unique ability to replicate as a linear plasmid with telomeres in its natural host E. coli during its prophage stage of life-cycle. To replicate as linear DNA, the virus produces a special enzyme called protelomerase that cleaves within its genome at a short recognition sequence known as the telomere occupancy site (tos) (Rybchin and Svarchevsky, 1999). Previous work in our lab have demonstrated that protelomerase-tos is able to assemble linear chromosome-like DNA containing the entire human β-globin genomic locus that can recapitulate accurate splicing of the globin mRNA when transferred into mammalian cells (Ooi et al., 2008). In parallel, we have developed enzymatic assays to verify functionality of protelomerase when expressed in bacteria (Chen and Narayanan, 2011) and mammalian cells (Chen and Narayanan, 2015; Chen et al., 2014). This PhD project will seek to further investigate the activity of the protelomerase enzyme expressed in the mammalian environment in order to gain insight for developing important technologies for genome engineering.
* Chen, Q. and Narayanan, K. (2015). Recombineering linear BACs. In Bacterial Artificial Chromosomes, 2nd Edition: Methods in Molecular Biology Series, K. Narayanan (Editor), Springer, New York.
* Chen, Q., Lee, C.W., Sim, E.U, and Narayanan, K. (2014) Induction of protein expression within E. coli bactofection vector for entry into mammalian cells. Human Gene Therapy Methods 25: 40-7.
* Chen, Q. and Narayanan, K. (2011). Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays. Analytical Biochemistry 414: 169-171.
* Ooi, Y.S., Warburton, P.E., Ravin, N.V., and Narayanan, K. (2008). Recombineering linear DNA that replicate stably in E. coli. Plasmid 59: 63-71.
* Rybchin, V.N., and Svarchevsky, A.N. (1999). The plasmid prophage N15: a linear DNA with covalently closed ends. Mol. Microbiol. 33: 895-903.
The candidate should hold a B.Sc (Hons) degree and MUST be a Malaysian citizen in order to be eligible for stipend support.Strong preference will be given to applicants with proven expertise in molecular biology/genetics who are highly motivated and proven to work independently with minimal supervision.
To apply, please email your CV and 2 letters of references (at least one must be from an immediate supervisor) explicitly addressing the criteria above. Only qualified candidates will be contacted to schedule for an interview.
For English requirements, please refer to this link: Doctor of Philosophy (PhD)