Difference between revisions of "Template:Heidelberg/project/standardization/Detection"

 
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                                   We developed this system further to minimize the background activity by combining the HRP DNAzyme with an F8 ribozyme.<x-ref>Wang2014</x-ref> This way the stem that avoids the formation of the G-quadruplex can be designed stronger, as the F8 DNAzyme is able to cleave of a part of this stem. This results into a weakening of the stem that inhibits HRP-mimicking DNAzyme activity and thus to the formation of the G-quadruplex. The F8 DNAzyme was fused with aptamers so that in the presence of the ligand the F8 DNAzyme becomes active which then activates the HRP-mimicking DNAzyme (Fig. 6).
 
                                   We developed this system further to minimize the background activity by combining the HRP DNAzyme with an F8 ribozyme.<x-ref>Wang2014</x-ref> This way the stem that avoids the formation of the G-quadruplex can be designed stronger, as the F8 DNAzyme is able to cleave of a part of this stem. This results into a weakening of the stem that inhibits HRP-mimicking DNAzyme activity and thus to the formation of the G-quadruplex. The F8 DNAzyme was fused with aptamers so that in the presence of the ligand the F8 DNAzyme becomes active which then activates the HRP-mimicking DNAzyme (Fig. 6).
 
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                        <h3 class="basicheader"> Outlook </h3>
 
 
<h3 class="subheader"> Further possibilities </h3>
 
<h3 class="subheader"> Further possibilities </h3>
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                                 <p class="basictext"> Similarly to the HRP DNAzyme other DNAzymes like the RNA-cleaving DNAzymes 7-18 and 10- 23 DNAzyme can be modified to become ligand dependent.<x-ref>Wanga2002</x-ref> Many more candidates like the ones described by Wang <i>et al.</i> can be applied for multiplexed detection of small molecules using our visualization method for western or southern blotting.   
 
                                 <p class="basictext"> Similarly to the HRP DNAzyme other DNAzymes like the RNA-cleaving DNAzymes 7-18 and 10- 23 DNAzyme can be modified to become ligand dependent.<x-ref>Wanga2002</x-ref> Many more candidates like the ones described by Wang <i>et al.</i> can be applied for multiplexed detection of small molecules using our visualization method for western or southern blotting.   
 
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Latest revision as of 02:46, 19 September 2015

HRP-based Detection

We developed this system further to minimize the background activity by combining the HRP DNAzyme with an F8 ribozyme.Wang2014 This way the stem that avoids the formation of the G-quadruplex can be designed stronger, as the F8 DNAzyme is able to cleave of a part of this stem. This results into a weakening of the stem that inhibits HRP-mimicking DNAzyme activity and thus to the formation of the G-quadruplex. The F8 DNAzyme was fused with aptamers so that in the presence of the ligand the F8 DNAzyme becomes active which then activates the HRP-mimicking DNAzyme (Fig. 6).

Further possibilities

Similarly to the HRP DNAzyme other DNAzymes like the RNA-cleaving DNAzymes 7-18 and 10- 23 DNAzyme can be modified to become ligand dependent.Wanga2002 Many more candidates like the ones described by Wang et al. can be applied for multiplexed detection of small molecules using our visualization method for western or southern blotting.

 

Figure 6. Low background switchable HRP-mimicking DNAzyme

The activity of the HRP-mimicking DNAzyme is controlled via a strong stem. A part of the stem is cleaved of by a switchable F8 DNAzyme in presence of a specific ligand.