I am a third year DPhil student in Dr Larin-Monaco's laboratory, and the main focus of my DPhil research is to develop gene expressing Human Artificial Chromosomes (HAC) in human embryonic stem cells (hES). The work on hES is in collaboration with Professor William James, at the Dunn School of Pathology. Human HAC are self-sufficient genetic structures that behave similar to endogenous chromosomes, capable of gene expression, replication and segregation during cell division. The use of HAC in gene therapy has great advantages over conventional gene therapy methods, as HAC do not require integration into the host genome and can encompass large DNA fragments in their construct. HAC also avoid problems with positional effects and silencing of the desired therapeutic gene. Thus, research on HAC is an exciting field with a very promising future in therapeutic medicine.
My goal is to utilize the Herpes Simplex Virus 1 amplicon technology developed by Richard Wade-Martins and Antonio Chiocca as a delivery system for HAC in hES cells. Thus far, I have established the conditions for efficient transduction of hES cells with HSV amplicons and stable clone selection. Furthermore, I have examined the viability and pluripotency of hES cells post HSV transduction, and established conditions for chromosome harvesting for cytogenetic analysis. Work is underway to isolate stable clones following transduction with HSV-1 HAC vectors.
MANDEGAR, M. A., MORALLI, D., KHOJA, S., COWLEY, S., CHAN, D. Y., YUSUF, M., MUKHERJEE, S., BLUNDELL, M. P., VOLPI, E. V., THRASHER, A. J., JAMES, W. & MONACO, Z. L. Functional human artificial chromosomes are generated and stably maintained in human embryonic stem cells. Hum Mol Genet, 20, 2905-13.
MORALLI, D., YUSUF, M., MANDEGAR, M. A., KHOJA, S., MONACO, Z. L. & VOLPI, E. V. An improved technique for chromosomal analysis of human ES and iPS cells. Stem Cell Rev, 7, 471-7.
GERSTEIN, A. C., CLEATHERO, L. A., MANDEGAR, M. A. & OTTO, S. P. Haploids adapt faster than diploids across a range of environments. J Evol Biol, 24, 531-40.
LEFEBVRE, L., MAR, L., BOGUTZ, A., OH-MCGINNIS, R., MANDEGAR, M. A., PADEROVA, J., GERTSENSTEIN, M., SQUIRE, J. A. & NAGY, A. (2009) The interval between Ins2 and Ascl2 is dispensable for imprinting centre function in the murine Beckwith-Wiedemann region. Hum Mol Genet, 18, 4255-67.
MANDEGAR, M. A. & OTTO, S. P. (2007) Mitotic recombination counteracts the benefits of genetic segregation. Proc Biol Sci, 274, 1301-7.
SARGENT, R. D., MANDEGAR, M. A. & OTTO, S. P. (2006) A model of the evolution of dichogamy incorporating sex-ratio selection, anther-stigma interference, and inbreeding depression. Evolution, 60, 934-44.
Keywords/ Research Area:
Human Embryonic Stem Cells, Gene Therapy, Human Artificial Chromosomes, Herpes Simplex (HSV).
EPA Cephalosporin Scholarship, University of Oxford
Post Graduate Doctoral Natural Sciences and Engineering Research Council of Canada (NSERC), Government of Canada