Skip to main content
SpringerLink
Log in

Session IV – Chromosome Engineering

Construction of neocentromere-based human minichromosomes for gene delivery and centromere studies

  • Research Article
  • Published:
Gene Therapy Submit manuscript

Abstract

Human neocentromeres are fully functional centromeres that arise naturally in non-centromeric regions devoid of α-satellite DNA. We have successfully produced a series of minichromosomes by telomere-associated truncation of a marker chromosome mardel(10) containing a neocentromere. The resulting minichromosomes are either linear or circular in nature, and range in size from approximately 650 kb to 2 Mb. These minichromosomes exhibit full centromeric activity, bind to essential centromere proteins, and are mitotically stable over many generations. They provide a useful system for dissecting the functional domains of complex eukaryotic centromeres and as vectors for therapeutic gene delivery.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2

Similar content being viewed by others

References

  1. Voullaire LE, Slater HR, Petrovic V, Choo KHA . A functional marker centromere with no detectable alpha-satellite, satellite III, or CENP-B protein: activation of a latent centromere? Am J Hum Genet 1993 52: 1153–1163

    CAS  PubMed  PubMed Central  Google Scholar 

  2. du Sart D et al. A functional neo-centromere formed through activation of a latent human centromere and consisting of non-alpha-satellite DNA Nat Genet 1997 16: 144–153

    Article  CAS  Google Scholar 

  3. Barry AE et al. Sequence analysis of an 80 kb human neocentromere Hum Mol Genet 1999 8: 217–227

    Article  CAS  Google Scholar 

  4. Barry AE et al. The 10q25 neocentromere and its inactive progenitor have identical primary nucleotide sequence: further evidence for epigenetic modification Genome Res 2000 10: 832–838

    Article  CAS  Google Scholar 

  5. Saffery R et al. Components of the human spindle checkpoint control mechanism localize specifically to the active centromere on dicentric chromosomes Hum Genet 2000 107: 376–384

    Article  CAS  Google Scholar 

  6. Saffery R et al. Human centromeres and neocentromeres show identical distribution patterns of >20 functionally important kinetochore-associated proteins Hum Mol Genet 2000 9: 175–185

    Article  CAS  Google Scholar 

  7. Lo AW et al. A 330 kb CENP-A binding domain and altered replication timing at a human neocentromere Embo J 2001 20: 2087–2096

    Article  CAS  Google Scholar 

  8. Lo AW et al. A novel chromatin immunoprecipitation and array (CIA) analysis identifies a 460-kb CENP-A-binding neocentromere DNA Genome Res 2001 11: 448–457

    Article  CAS  Google Scholar 

  9. Choo KHA . Domain organisation at the centromere and neocentromere Dev Cell 2001 1: 165–177

    Article  CAS  Google Scholar 

  10. Warburton PE et al. Molecular cytogenetic analysis of eight inversion duplications of human chromosome 13q that each contain a neocentromere Am J Hum Genet 2000 66: 1794–1806

    Article  CAS  Google Scholar 

  11. Farr CJ et al. Generation of a human X-derived minichromosome using telomere-associated chromosome fragmentation Embo J 1995 14: 5444–5454

    Article  CAS  Google Scholar 

  12. Heller R, Brown KE, Burgtorf C . Brown WR.. Mini-chromosomes derived from the human Y chromosome by telomere directed chromosome breakage Proc Natl Acad Sci USA 1996 93: 7125–7130

    Article  CAS  Google Scholar 

  13. Saffery R et al. Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation Proc Natl Acad Sci USA 2001 98: 5705–5710

    Article  CAS  Google Scholar 

  14. Brown WRA, Mee PJ, Shen MH . Artificial chromosomes: ideal vectors? Trends Biotech 2000 18: 218–223

    Article  CAS  Google Scholar 

  15. Willard HF . Neocentromeres and human artificial chromosomes: an unnatural act Proc Natl Acad Sci USA 2001 98: 5705–5710

    Article  Google Scholar 

  16. Choo KHA . Engineering human chromosomes for gene therapy studies Trends Mol Med 2001 7: 235–237

    Article  CAS  Google Scholar 

  17. Saffery R, Choo KHA . Strategies for engineering human chromosomes with therapeutic potential J Gene Med 2002 4: 5–13

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia

    L H Wong, R Saffery & K H A Choo

Authors
  1. L H Wong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R Saffery
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K H A Choo
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wong, L., Saffery, R. & Choo, K. Construction of neocentromere-based human minichromosomes for gene delivery and centromere studies. Gene Ther 9, 724–726 (2002). https://doi.org/10.1038/sj.gt.3301756

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301756

  • Springer Nature Limited

Keywords

This article is cited by

Search

Navigation