Development and Study of Flavone Derivatives for Triplex Binding
Poster Number
5C
Introduction/Abstract
Triplex DNA forms upon the sequence-specific binding of a triplex-forming oligonucleotide (TFO) into the major groove of a duplex DNA. Triplexes that form endogenously are known as H-DNA, and they can be found in promoter regions, telomerases, and spliceosome complexes. They are also implicated in genetic diseases such as Friedreich’s ataxia. Furthermore, certain helicases have been found to unwind triplex DNA, highlighting the physiological relevance of triplex DNA. Because of its high specificity, triplex DNA makes an ideal target for antigene therapy. Upon the formation of triplex DNA, normal cellular machinery can no longer recognize and bind to that region of the genome, which can lead to a complete halt of the synthesis of that particular protein. Unfortunately, triplex DNA is less stable than duplex DNA under physiological conditions, so there needs to be other ways to help stabilize the triplex DNA.
Purpose
Our group has discovered a class of flavonoid derivatives that can strongly stabilize triplex DNA but do not have an effect on duplex DNA. In the poster, we report further structural variations of flavonoid derivatives in order to determine what moieties contribute the most to stability. We also studied the effect of triplex DNA inhibiting restriction enzyme activity. Through these studies, we hope to see the new compounds bind specifically to triplexes and that the formation of triplex DNA can inhibit restriction enzyme activity.
Method
Novel flavonoid derivatives were synthesized and subject to thermal denaturation monitored by UV to test for stability. Restriction enzyme digestion assays were performed to see if triplex formation can inhibit restriction enzyme activity.
Results
The synthesized compounds show an increased melting temperature of the triplex DNA while having no effect on duplex DNA. The side chain at the 5-position is important for triplex binding. Furthermore, the restriction enzyme digestion assays show inhibition of restriction enzyme activity.
Significance
We have successfully developed a class of novel triplex-specific binding flavonoid derivatives.
Location
Library and Learning Center, 3601 Pacific Ave., Stockton, CA 95211
Format
Poster Presentation
Development and Study of Flavone Derivatives for Triplex Binding
Library and Learning Center, 3601 Pacific Ave., Stockton, CA 95211
Triplex DNA forms upon the sequence-specific binding of a triplex-forming oligonucleotide (TFO) into the major groove of a duplex DNA. Triplexes that form endogenously are known as H-DNA, and they can be found in promoter regions, telomerases, and spliceosome complexes. They are also implicated in genetic diseases such as Friedreich’s ataxia. Furthermore, certain helicases have been found to unwind triplex DNA, highlighting the physiological relevance of triplex DNA. Because of its high specificity, triplex DNA makes an ideal target for antigene therapy. Upon the formation of triplex DNA, normal cellular machinery can no longer recognize and bind to that region of the genome, which can lead to a complete halt of the synthesis of that particular protein. Unfortunately, triplex DNA is less stable than duplex DNA under physiological conditions, so there needs to be other ways to help stabilize the triplex DNA.
