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© 1994 Oxford University Press

OTHER

Addition of functional human telomeres to YACs

Stephen S. Taylor* and Chris Tyler Smith

CRC Chromosome Molecular Biology Group, Department of Biochemistry, University of Oxford South Parks Road, Oxford OX1 3QU, UK

*To whom correspondence should be addressed

Received April 22, 1994; Accepted June 13, 1994

Linear mammalian artificial chromosomes (MACs) will require functional telomeres, a centromere and the abllity to repllcate autonomously. We are investigating the possibility of developing MACs from yeast artificial chromosomes (YACs). Retrofitting vectors have been constructed to replace YAC telomeres with cloned human telomeric DNA. A modified YAC was introduced into mammalian cells by spheroplast tusion and the frequency with which the retrofitted human telomeric DNA seeded the formation of a new telomere was determined by Bal31 digestion and cytogenetic analysis. The telomere adjacent to the selectable marker gene was functional in 5/46 clones (11%) while the telomere 200 kb away at the other end of the YAC was functional in 1/46 clones (2%). These results indicate that despite the in vivo modification of the end of the telomere by the addition of yeast sequences, human telomeres will function at a high enough frequency to allow the construction of MACs by this route.


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