Site-Specific Cleavage of a Yeast Chromosome by Oligonucleotide-Directed Triple-Helix Formation
- Creators
- Strobel, Scott A.
-
Dervan, Peter B.
Abstract
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.
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.Additional details
- Eprint ID
- 54063
- DOI
- 10.1126/science.2195655
- Resolver ID
- CaltechAUTHORS:20150126-091403335
- GM 42966
- NIH
- Created
-
2015-01-26Created from EPrint's datestamp field
- Updated
-
2021-11-10Created from EPrint's last_modified field