Addition of functional human telomeres to YACs.

Linear mammalian artificial chromosomes (MACs) will require functional telomeres, a centromere and the ability to replicate autonomously. We are investigating the possibility of developing MACs from yeast artificial chromosomes (YACs). Retrofitting vectors have been constructed to replace YAC telome...

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Bibliografski detalji
Glavni autori: Taylor, S, Larin, Z, Smith, C
Format: Journal article
Jezik:English
Izdano: 1994
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author Taylor, S
Larin, Z
Smith, C
author_facet Taylor, S
Larin, Z
Smith, C
author_sort Taylor, S
collection OXFORD
description Linear mammalian artificial chromosomes (MACs) will require functional telomeres, a centromere and the ability to replicate 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 fusion 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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spelling oxford-uuid:9f1516f5-8f6d-4e9a-ac69-eaee6de251ef2022-03-27T00:54:47ZAddition of functional human telomeres to YACs.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9f1516f5-8f6d-4e9a-ac69-eaee6de251efEnglishSymplectic Elements at Oxford1994Taylor, SLarin, ZSmith, CLinear mammalian artificial chromosomes (MACs) will require functional telomeres, a centromere and the ability to replicate 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 fusion 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.
spellingShingle Taylor, S
Larin, Z
Smith, C
Addition of functional human telomeres to YACs.
title Addition of functional human telomeres to YACs.
title_full Addition of functional human telomeres to YACs.
title_fullStr Addition of functional human telomeres to YACs.
title_full_unstemmed Addition of functional human telomeres to YACs.
title_short Addition of functional human telomeres to YACs.
title_sort addition of functional human telomeres to yacs
work_keys_str_mv AT taylors additionoffunctionalhumantelomerestoyacs
AT larinz additionoffunctionalhumantelomerestoyacs
AT smithc additionoffunctionalhumantelomerestoyacs