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Session IV – Chromosome Engineering

Retrofitting of a satellite repeat DNA-based murine artificial chromosome (ACes) to contain loxP recombination sites

Gene Therapy volume 9, pages 719–723 (2002)Cite this article

Abstract

A satellite DNA-based mammalian artificial chromosome (ACes) was generated and subsequently modified by targeting of a loxP-red fluorescent protein (RFP) expression cassette via homologous recombination into a ribosomal DNA (rDNA)-containing locus. Clones containing correctly targeted ACes were identified by PCR from populations of RFP-expressing cells enriched by FACS sorting and were further characterized by fluorescent in situ hybridization. The targeted ACes maintained its ability to be purified to near homogeneity. Studies are currently underway to further characterize the functionality, carrying capacity, stability and transfectability of this modified ACes.

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Acknowledgements

We would like to thank Drs Gyula Hadlaczky, Amy Greene, Joseph Zendegui, Ross Durland and Brenda Grimes for their helpful comments and suggestions. The expert technical assistance of Spencer Watson, Diane Monteith, Josephine Leung and Tom Stodola is also gratefully acknowledged.

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Authors and Affiliations

  1. Chromos Molecular Systems Inc., Burnaby, BC, Canada

    S Stewart, N MacDonald, E Perkins, G deJong, C Perez & M Lindenbaum

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  1. S Stewart
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  2. N MacDonald
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  3. E Perkins
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  4. G deJong
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  5. C Perez
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Stewart, S., MacDonald, N., Perkins, E. et al. Retrofitting of a satellite repeat DNA-based murine artificial chromosome (ACes) to contain loxP recombination sites. Gene Ther 9, 719–723 (2002). https://doi.org/10.1038/sj.gt.3301757

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