A high-throughput screen for detection of compound-dependent phosphodiester bond cleavage at abasic sites.
Anal Biochem
; 513: 93-97, 2016 11 15.
Article
in En
| MEDLINE
| ID: mdl-27594348
ABSTRACT
We have employed a DNA molecular beacon with a real abasic site, namely a 2-deoxyribose, in a fluorescent high-throughput assay to identify artificial nucleases that cleave at abasic sites. We screened a 1280 compound chemical library and identified a compound that functions as an artificial nuclease. We validated a key structure-activity relationship necessary for abasic site cleavage using available analogs of the identified artificial nuclease. We also addressed the activity of the identified compound with dose titrations in the absence and presence of a source of non-specific DNA. Finally, we characterized the phosphodiester backbone cleavage at the abasic site using denaturing gel electrophoresis. This study provides a useful template for researchers seeking to rapidly identify new artificial nucleases.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
DNA
/
Deoxyribonucleases
/
Deoxyribose
Type of study:
Diagnostic_studies
Language:
En
Year:
2016
Type:
Article