α-satellite DNA and vector composition influence rates of human artificial chromosome formation

Brenda Grimes, Angela A. Rhoades, Huntington F. Willard

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Human artificial chromosomes (HACs) have been proposed as a new class of potential gene transfer and gene therapy vector. HACs can be formed when bacterial cloning vectors containing α-satellite DNA are transfected into cultured human cells. We have compared the HAC-forming potential of different sequences to identify features critical to the efficiency of the process. Chromosome 17 or 21 α-satellite arrays are highly competent HAC-forming substrates in this assay. In contrast, a Y-chromosome-derived α-satellite sequence is inefficient, suggesting that centromere specification is at least partly dependent on DNA sequence. The length of the input array is also an important determinant, as reduction of the chromosome-17-based array from 80 kb to 35 kb reduced the frequency of HAC formation. In addition to the α-satellite component, vector composition also influenced HAC formation rates, size, and copy number. The data presented here have a significant impact on the design of future HAC vectors that have potential to be developed for therapeutic applications and as tools for investigating human chromosome structure and function.

Original languageEnglish (US)
Pages (from-to)798-805
Number of pages8
JournalMolecular Therapy
Volume5
Issue number6
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Human Artificial Chromosomes
Satellite DNA
Chromosomes, Human, Pair 17
Chromosome Structures
Chromosomes, Human, Pair 21
Genetic Vectors
Centromere
Y Chromosome
Human Chromosomes
Genetic Therapy
Cultured Cells

Keywords

  • α-satellite DNA
  • Bacterial cloning vectors
  • Human artificial chromosome

ASJC Scopus subject areas

  • Molecular Biology

Cite this

α-satellite DNA and vector composition influence rates of human artificial chromosome formation. / Grimes, Brenda; Rhoades, Angela A.; Willard, Huntington F.

In: Molecular Therapy, Vol. 5, No. 6, 2002, p. 798-805.

Research output: Contribution to journalArticle

Grimes, Brenda ; Rhoades, Angela A. ; Willard, Huntington F. / α-satellite DNA and vector composition influence rates of human artificial chromosome formation. In: Molecular Therapy. 2002 ; Vol. 5, No. 6. pp. 798-805.
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