David Y.L. Chan

DPhil Student

david

Research Summary:

Human artificial chromosomes, based on the human centromeric alpha satellite DNA, have been successfully transferred to murine cells using the microcell mediated chromosome transfer technique (MMCT). However, this procedure may induce rearrangement in the HAC structure, that can affect its stability, or the expression of the genes carried by the HAC.  As a result, MMCT is not ideal for gene therapy purposes. The direct generation of artificial chromosomes (de novo) in murine cells is a tempting approach for gene therapy in murine models of genetic deficiencies. Using human alpha satellite to seed HAC formation in murine cells has a very poor efficiency, and does not work in all cell types.  I am now isolating and cloning mouse centromeric sequences for the direct generation of mouse artificial chromosomes (MAC) in murine embryonic stell cells and fibroblast cell lines using HSV-1 amplicon delivery system.

My preliminary results showed that large stretches of mouse major satellites are capable of triggering the formation of extrachromosomal structures, albeit at an extremely low efficiency. I am now focusing on increasing the efficiency/stability of MAC formation.

MACs

Publication List:

Moralli,D., Chan,D.Y., Jefferson,A., Volpi,E.V., and Monaco,Z.L. (2009). HAC stability in murine cells is influenced by nuclear localization and chromatin organization. BMC. Cell Biol. 10, 18.

Chan,D.Y., Chen,G.G., Poon,W.S., and Liu,P.C. (2008). Lovastatin sensitized human glioblastoma cells to TRAIL-induced apoptosis. J. Neurooncol. 86, 273-283.

Ho PW, Chan DY, Kwok KH, Chu AC, Ho JW, Kung MH, Ramsden DB, Ho SL (2005) Methyl-4-phenylpyridinium ion modulates expression of mitochondrial uncoupling proteins 2, 4, and 5 in catecholaminergic (SK-N-SH) cells. J Neurosci Res 81: 261-268.

David Y. L. Chan, Wai Keung Leung, George Chen and Wai Sun Poon; Tumor necrosis factor related apoptosis-inducing ligand modulate mRNA expression profile in human glioma cell line A172; The 96th Annual Meeting 2005, American Association for Cancer Research.

De novo mouse artificial chromosome formation.

Keywords:

Murine Artificial Chromosomes (MACs), mouse satellites, mouse embryonic stem cells.

Contact Details

Email: ylchan@well.ox.ac.uk

Sponsors:

Clarendon Award (Oxford), China/Hong Kong Oxford Scholarship.