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Mix and measure fluorescence screening for selective quadruplex binders.

Paramasivan S, Bolton PH - Nucleic Acids Res. (2008)

Bottom Line: The successful targeting of a particular quadruplex structure requires distinguishing that structure from all of the other quadruplex structures that may be present.The screening is based on observing the increase or decrease in the fluorescence of the reporter molecules.The selectivity of a set of test molecules has been determined by this approach.

View Article: PubMed Central - PubMed

Affiliation: Chemistry Department, Wesleyan University, Middletown, CT 06459, USA.

ABSTRACT
The human genome contains thousands of regions, including that of the telomere, that have the potential to form quadruplex structures. Many of these regions are potential targets for therapeutic intervention. There are many different folding patterns for quadruplex DNAs and the loops exhibit much more variation than do the quartets. The successful targeting of a particular quadruplex structure requires distinguishing that structure from all of the other quadruplex structures that may be present. A mix and measure fluorescent screening method has been developed, that utilizes multiple reporter molecules that bind to different features of quadruplex DNA. The reporter molecules are used in combination with DNAs that have a variety of quadruplex structures. The screening is based on observing the increase or decrease in the fluorescence of the reporter molecules. The selectivity of a set of test molecules has been determined by this approach.

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The results of the screening are shown for all of the DNAs and all three reporter molecules. Increases in fluorescence are in blue and decreases are shown in red with the darker shades indicating a larger effect as indicated in the figure. The color chart representation is used to allow for visual comparison of the patterns obtained with different test molecules.
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Figure 6: The results of the screening are shown for all of the DNAs and all three reporter molecules. Increases in fluorescence are in blue and decreases are shown in red with the darker shades indicating a larger effect as indicated in the figure. The color chart representation is used to allow for visual comparison of the patterns obtained with different test molecules.

Mentions: The screening was carried out using a conventional microplate reader. The fluorescence of the DNA–reporter molecule samples was measured in the presence and in the absence of the test ligand. The results are presented in Figure 6 with red indicating a decrease in fluorescence and blue an increase with the darker shades indicating a larger change. The color chart mode of presentation allows visual comparison of the results obtained for the test ligands.Figure 6.


Mix and measure fluorescence screening for selective quadruplex binders.

Paramasivan S, Bolton PH - Nucleic Acids Res. (2008)

The results of the screening are shown for all of the DNAs and all three reporter molecules. Increases in fluorescence are in blue and decreases are shown in red with the darker shades indicating a larger effect as indicated in the figure. The color chart representation is used to allow for visual comparison of the patterns obtained with different test molecules.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2553591&req=5

Figure 6: The results of the screening are shown for all of the DNAs and all three reporter molecules. Increases in fluorescence are in blue and decreases are shown in red with the darker shades indicating a larger effect as indicated in the figure. The color chart representation is used to allow for visual comparison of the patterns obtained with different test molecules.
Mentions: The screening was carried out using a conventional microplate reader. The fluorescence of the DNA–reporter molecule samples was measured in the presence and in the absence of the test ligand. The results are presented in Figure 6 with red indicating a decrease in fluorescence and blue an increase with the darker shades indicating a larger change. The color chart mode of presentation allows visual comparison of the results obtained for the test ligands.Figure 6.

Bottom Line: The successful targeting of a particular quadruplex structure requires distinguishing that structure from all of the other quadruplex structures that may be present.The screening is based on observing the increase or decrease in the fluorescence of the reporter molecules.The selectivity of a set of test molecules has been determined by this approach.

View Article: PubMed Central - PubMed

Affiliation: Chemistry Department, Wesleyan University, Middletown, CT 06459, USA.

ABSTRACT
The human genome contains thousands of regions, including that of the telomere, that have the potential to form quadruplex structures. Many of these regions are potential targets for therapeutic intervention. There are many different folding patterns for quadruplex DNAs and the loops exhibit much more variation than do the quartets. The successful targeting of a particular quadruplex structure requires distinguishing that structure from all of the other quadruplex structures that may be present. A mix and measure fluorescent screening method has been developed, that utilizes multiple reporter molecules that bind to different features of quadruplex DNA. The reporter molecules are used in combination with DNAs that have a variety of quadruplex structures. The screening is based on observing the increase or decrease in the fluorescence of the reporter molecules. The selectivity of a set of test molecules has been determined by this approach.

Show MeSH