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...
| Glavni autori: | , , |
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| Format: | Journal article |
| Jezik: | English |
| Izdano: |
1994
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| _version_ | 1826287939991109632 |
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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. |
| first_indexed | 2024-03-07T02:06:12Z |
| format | Journal article |
| id | oxford-uuid:9f1516f5-8f6d-4e9a-ac69-eaee6de251ef |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:06:12Z |
| publishDate | 1994 |
| record_format | dspace |
| 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 |