Site-specific cleavage of human chromosome 4 mediated by triple-helix formation
Abstract
Direct physical isolation of specific DNA segments from the human genome is a necessary goal in human genetics. For testing whether triple-helix mediated enzymatic cleavage can liberate a specific segment of a human chromosome, the tip of human chromosome 4, which contains the entire candidate region for the Huntington's disease gene, was chosen as a target. A 16-base pyrimidine oligodeoxyribonucleotide was able to locate a 16-base pair purine target site within more than 10 gigabase pairs of genomic DNA and mediate the exact enzymatic cleavage at that site in more than 80 percent yield. The recognition motif is sufficiently generalizable that most cosmids should contain a sequence targetable by triple-helix formation. This method may facilitate the orchestrated dissection of human chromosomes from normal and affected individuals into megabase sized fragments and facilitate the isolation of candidate gene loci.
Additional Information
© 1991 American Association for the Advancement of Science. 3 September 1991; Accepted 12 November 1991. We thank J. F. Gusella for cosmid 8C10I5, and assistance with the physical map location of the target site; the members of the Hereditary Disease Foundation HD Collaborative Research group for STS-PCR primer sequences and helpful discussions; the Howard Hughes Medical Institute for a predoctoral fellowship to S.A.S.; the Joan and William Schreyer Research Fund for postdoctoral fellowship to L.D.S.; the National Institutes of Health (HG00098 and HG003293 and the Hereditary Disease Foundation for grant support.Additional details
- Eprint ID
- 53876
- DOI
- 10.1126/science.1836279
- Resolver ID
- CaltechAUTHORS:20150120-111915277
- Howard Hughes Medical Institute (HHMI)
- Joan and William Schreyer Reseach Fund
- HG00098
- NIH
- HG00329
- NIH
- Hereditary Disease Foundation
- Created
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2015-01-20Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field