Site-specific cleavage of human chromosome 4 mediated by triple-helix formation

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 regio...

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Published in:Science (American Association for the Advancement of Science) 1991-12, Vol.254 (5038), p.1639-1642
Main Authors: Strobel, S A, Doucette-Stamm, L A, Riba, L, Housman, D E, Dervan, P B
Format: Article
Language:English
Subjects:
Base Sequence
Cellular biology
Chromosome Mapping - methods
Chromosomes, Human, Pair 4 - ultrastructure
DNA
DNA Damage
Enzymes
Genetics
genomes
Humans
Huntington Disease - genetics
Hydrogen Bonding
Molecular Sequence Data
Nucleic Acid Conformation
Oligodeoxyribonucleotides - chemistry
Restriction Mapping
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description 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.
doi_str_mv 10.1126/science.1836279
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subjects Base Sequence
Cellular biology
Chromosome Mapping - methods
Chromosomes, Human, Pair 4 - ultrastructure
DNA
DNA Damage
Enzymes
Genetics
genomes
Humans
Huntington Disease - genetics
Hydrogen Bonding
Molecular Sequence Data
Nucleic Acid Conformation
Oligodeoxyribonucleotides - chemistry
Restriction Mapping
title Site-specific cleavage of human chromosome 4 mediated by triple-helix formation
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