Efficiency of de novo centromere formation in human artificial chromosomes.
In a comparative study, we show that human artificial chromosome (HAC) vectors based on alpha-satellite (alphoid) DNA from chromosome 17 but not the Y chromosome regularly form HACs in HT1080 human cells. We constructed four structurally similar HAC vectors, two with chromosome 17 or Y alphoid DNA (...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
2002
|
| _version_ | 1826274935618666496 |
|---|---|
| author | Mejía, J Alazami, A Willmott, A Marschall, P Levy, E Earnshaw, W Larin, Z |
| author_facet | Mejía, J Alazami, A Willmott, A Marschall, P Levy, E Earnshaw, W Larin, Z |
| author_sort | Mejía, J |
| collection | OXFORD |
| description | In a comparative study, we show that human artificial chromosome (HAC) vectors based on alpha-satellite (alphoid) DNA from chromosome 17 but not the Y chromosome regularly form HACs in HT1080 human cells. We constructed four structurally similar HAC vectors, two with chromosome 17 or Y alphoid DNA (17alpha, Yalpha) and two with 17alpha or Yalpha and the hypoxanthine guanine phosphoribosyltransferase locus (HPRT1). The 17alpha HAC vectors generated artificial minichromosomes in 32-79% of the HT1080 clones screened, compared with only approximately 4% for the Yalpha HAC vectors, indicating that Yalpha is inefficient at forming a de novo centromere. The 17alpha HAC vectors produced megabase-sized, circular HACs containing multiple copies of alphoid fragments (60-250 kb) interspersed with either vector or HPRT1 DNA. The 17alpha-HPRT1 HACs were less stable than those with 17alpha only, and these results may influence the design of new HAC gene transfer vectors. |
| first_indexed | 2024-03-06T22:51:03Z |
| format | Journal article |
| id | oxford-uuid:5ec92bef-b285-49d4-88ab-4550ef83c4ac |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:51:03Z |
| publishDate | 2002 |
| record_format | dspace |
| spelling | oxford-uuid:5ec92bef-b285-49d4-88ab-4550ef83c4ac2022-03-26T17:42:50ZEfficiency of de novo centromere formation in human artificial chromosomes.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:5ec92bef-b285-49d4-88ab-4550ef83c4acEnglishSymplectic Elements at Oxford2002Mejía, JAlazami, AWillmott, AMarschall, PLevy, EEarnshaw, WLarin, ZIn a comparative study, we show that human artificial chromosome (HAC) vectors based on alpha-satellite (alphoid) DNA from chromosome 17 but not the Y chromosome regularly form HACs in HT1080 human cells. We constructed four structurally similar HAC vectors, two with chromosome 17 or Y alphoid DNA (17alpha, Yalpha) and two with 17alpha or Yalpha and the hypoxanthine guanine phosphoribosyltransferase locus (HPRT1). The 17alpha HAC vectors generated artificial minichromosomes in 32-79% of the HT1080 clones screened, compared with only approximately 4% for the Yalpha HAC vectors, indicating that Yalpha is inefficient at forming a de novo centromere. The 17alpha HAC vectors produced megabase-sized, circular HACs containing multiple copies of alphoid fragments (60-250 kb) interspersed with either vector or HPRT1 DNA. The 17alpha-HPRT1 HACs were less stable than those with 17alpha only, and these results may influence the design of new HAC gene transfer vectors. |
| spellingShingle | Mejía, J Alazami, A Willmott, A Marschall, P Levy, E Earnshaw, W Larin, Z Efficiency of de novo centromere formation in human artificial chromosomes. |
| title | Efficiency of de novo centromere formation in human artificial chromosomes. |
| title_full | Efficiency of de novo centromere formation in human artificial chromosomes. |
| title_fullStr | Efficiency of de novo centromere formation in human artificial chromosomes. |
| title_full_unstemmed | Efficiency of de novo centromere formation in human artificial chromosomes. |
| title_short | Efficiency of de novo centromere formation in human artificial chromosomes. |
| title_sort | efficiency of de novo centromere formation in human artificial chromosomes |
| work_keys_str_mv | AT mejiaj efficiencyofdenovocentromereformationinhumanartificialchromosomes AT alazamia efficiencyofdenovocentromereformationinhumanartificialchromosomes AT willmotta efficiencyofdenovocentromereformationinhumanartificialchromosomes AT marschallp efficiencyofdenovocentromereformationinhumanartificialchromosomes AT levye efficiencyofdenovocentromereformationinhumanartificialchromosomes AT earnshaww efficiencyofdenovocentromereformationinhumanartificialchromosomes AT larinz efficiencyofdenovocentromereformationinhumanartificialchromosomes |