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Site-Specific Cleavage of a Yeast Chromosome by Oligonucleotide-Directed Triple-Helix Formation

Strobel, Scott A. and Dervan, Peter B. (1990) Site-Specific Cleavage of a Yeast Chromosome by Oligonucleotide-Directed Triple-Helix Formation. Science, 249 (4964). pp. 73-75. ISSN 0036-8075. doi:10.1126/science.2195655.

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Oligonucleotides equipped with EDTA•Fe can bind specifically to duplex DNA by triple-helix formation and produce double-strand cleavage at binding sites greater than 12 base pair in size. To demonstrate that oligonucleotide-directed triple-helix formation is a viable chemical approach for the site-specific cleavage of large genomic DNA, an oligonucleotide with EDTA•Fe at the 5' and 3' ends was targeted to a 20-base pair sequence in the 340-kilobase pair cromosome III of Saccharomyces cerevisiae. Double-trand cleavage products of the correct size and location were observed, indicating that the oligonucleotide bound and cleaved the target site among almost 14 megabase pairs of DNA. Because oligonucleotide-directed triple-helix formation has the potential to be a general solution for DNA recognition, this result has implications for physical mapping of chromosomes.

Item Type:Article
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Dervan, Peter B.0000-0001-8852-7306
Additional Information:© 1990 American Association for the Advancement of Science. 24 January 1990; accepted 1 May 1990. We thank S. Emr for yeast strain SEY6210, the shuttle vector YEp503, and assistance in the design of the yeast transformation; R. Schekman for the HIS4 marker in YCp503; J. Campbell for the PGK1 promoter on pMA91; B. Birren for yeast strain YNN295 and assistance with pulsed-field gel electrophoresis; and the Howard Hughes Institute for a predoctoral fellowship to S.A.S. Supported by NIH grant GM 42966.
Funding AgencyGrant Number
NIHGM 42966
Issue or Number:4964
Record Number:CaltechAUTHORS:20150126-091403335
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Official Citation:Site-Specific Cleavage of a Yeast Chromosome by Oligonucleotide-Directed Triple-Helix Formation Scott A. Strobel and Peter B. Dervan Science New Series, Vol. 249, No. 4964 (Jul. 6, 1990) , pp. 73-75 Published by: American Association for the Advancement of Science Stable URL:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:54063
Deposited By: Ruth Sustaita
Deposited On:26 Jan 2015 17:53
Last Modified:10 Nov 2021 20:28

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