Triple helix-mediated single-site enzymatic cleavage of megabase genomic DNA
- Creators
- Strobel, S. A.
-
Dervan, P. B.
Abstract
This chapter explains the triple helix-mediated single-site enzymatic cleavage of megabase genomic DNA. Oligonucleotide-directed triple helix formation is a generalizable chemical approach for the recognition and cleavage of a single target site within several megabase pairs of duplex genomic DNA. The generalizability of triple helix-mediated enzymatic cleavage affords high specificity that can be readily customized to unique genetic markers without artificial insertion of a target sequence. Extensive sequencing to identify target sites could be avoided by using degenerate oligonucleotides to screen genetic markers for overlapping triple helix/ endonuclease sites. The potential generalizability of triple helix-mediated Achilles' heel cleavage, a technique capable of near-quantitative cleavage at a single site in at least 14 megabase pairs of DNA, could assist in physical mapping of chromosomal DNA and expedite isolation of DNA segments linked to the disease.
Additional Information
© 1992 Elsevier Inc. We are grateful to Dr. John Hanish for helpful discussions, the National Institutes of Health for grant support, and the Howard Hughes Medical Institute for a predoctoral fellowship to S.A.S.Additional details
- Eprint ID
- 66952
- Resolver ID
- CaltechAUTHORS:20160510-145809040
- NIH
- Howard Hughes Medical Institute (HHMI)
- Created
-
2016-05-19Created from EPrint's datestamp field
- Updated
-
2021-11-11Created from EPrint's last_modified field
- Series Name
- Methods in Enzymology
- Series Volume or Issue Number
- 216