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<li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Team" onmouseover="mopen('m2')" onmouseout="mclosetime()" style="padding:15px 0px;">Team</a>
<li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Team" onmouseover="mopen('m2')" onmouseout="mclosetime()" style="padding:15px 0px;">Team</a>
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<a href="#">Item1</a>
 
<a href="#">Item2</a>
<a href="#">Item3</a>
<a href="#">Item4</a>
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<li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs" onmouseover="mopen('m3')" onmouseout="mclosetime()" style="padding:15px 0px;">Designs</a>
<li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs" onmouseover="mopen('m3')" onmouseout="mclosetime()" style="padding:15px 0px;">Designs</a>
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                <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs">Background</a>
<a href="#">Item2</a>
<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/Self-assemblyTrack">Self-assembly Track</a>
<a href="#">Item3</a>
<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/DeliveryDevice">The Delivery Device</a>
<a href="#">Item4</a>
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<li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results" onmouseover="mopen('m4')" onmouseout="mclosetime()" style="height:35px;padding:8px 0px 7px 0px;">Experiments &amp; Results</a>
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<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results">Polymerizing</a>
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<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results/DeliveryDevice">Delivery Device</a>
<a href="#">Item4</a>
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     <li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Attributions" onmouseover="mopen('m6')" onmouseout="mclosetime()" style="padding:15px 0px;">Attributions</a>
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<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs" style="font-size:14px;"><b>Background</b></a>
<p>HCR、DNAzyme、DNA-walker、Origami</p>
<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/Self-assemblyTrack" style="font-size:14px;"><b>Self-assembly Track</b></a>
<p>T1-T4、The polymerization reaction</p>
<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/DeliveryDevice"><b>The Delivery device</b></a>
<br/><br/>
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<span style="color:#000;font-size:22px;"><b>Background</b></span>
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<table id="toc" class="toc" summary="Contents" style="font-family:Calibri;float:right;font-size:120%;"><tbody><tr><td><div id="toctitle"><h2>Contents</h2></div>
<table id="toc" class="toc" summary="Contents" style="font-family:Calibri;float:right;font-size:120%;"><tbody><tr><td><div id="toctitle"><h2>Contents</h2></div>
<ul>
<ul>
<li class="toclevel-1"><a href="#The_DNA.E2.80.94Track"><span class="tocnumber">1</span> <span class="toctext">The DNA—Track</span></a></li>
<li class="toclevel-1"><a href="#HCR"><span class="tocnumber">1</span> <span class="toctext">HCR</span></a></li>
<li class="toclevel-1"><a href="#The_Walker"><span class="tocnumber">2</span> <span class="toctext">The Walker</span></a></li>
<li class="toclevel-1"><a href="#DNAzyme"><span class="tocnumber">2</span> <span class="toctext">DNAzyme</span></a></li>
<li class="toclevel-1"><a href="#Agarose_electrophoresis"><span class="tocnumber">3</span> <span class="toctext">Agarose electrophoresis</span></a></li>
<li class="toclevel-1"><a href="#DNA-walker"><span class="tocnumber">3</span> <span class="toctext">DNA-walker</span></a></li>
<li class="toclevel-1"><a href="#PAGE"><span class="tocnumber">4</span> <span class="toctext">PAGE</span></a>
<li class="toclevel-1"><a href="#Origami"><span class="tocnumber">4</span> <span class="toctext">Origami</span></a>
<ul>
<li class="toclevel-2"><a href="#Materials"><span class="tocnumber">4.1</span> <span class="toctext">Materials</span></a></li>
<li class="toclevel-2"><a href="#Methods"><span class="tocnumber">4.2</span> <span class="toctext">Methods</span></a></li>
</ul>
</li>
<li class="toclevel-1"><a href="#PCR_Annealing_Procedure_.28for_Logic_Gate.29"><span class="tocnumber">5</span> <span class="toctext">PCR Annealing Procedure (for Logic Gate)</span></a></li>
<li class="toclevel-1"><a href="#Recipe"><span class="tocnumber">6</span> <span class="toctext">Recipe</span></a></li>
</ul>
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__NOTOC__
__NOTOC__
=The DNA—Track =
= HCR =


Stable DNA monomers assemble only upon exposure to atarget DNA fragment. In the simplest version of this process, two stable species of DNA hairpins coexist in solution until the introduction of initiator strands triggers a cascade of hybridization events that yields nicked double helices analogous to alternating copolymers. The average molecular weight of the HCR products varies inversely with initiator concentration. Amplification of more diverse recognition events can be achieved by coupling HCR to aptamer triggers. This functionality allows DNA to act as an amplifying transducer for biosensing applications.


* Version 1
[[Image:TJU-figure2.1.1.png|thumb|600px|center|'''Figure2.1.1 (a–c)''' Secondary structure schematic of HCR function. Letters marked with * are complementary to the corresponding unmarked letter.(a) Hairpins H1 and H2 are stable in the absence of initiator I. (b) I nucleates at the sticky end of H1 and undergoes an unbiased strand displacement interaction to open the hairpin. (c) The newly exposed sticky end of H1 nucleates at the sticky end of H2 and opens the hairpin to expose a sticky end on H2 that is identical in sequence to I. Hence, each copy of I can propagate a chain reaction of hybridization events between alternating H1 and H2 hairpins to form a nicked double-helix, amplifying the signal of initiator binding. Figure and introduction from <b>Robert M. Dirks and Niles A. Pierce</b> <sub>PNAS  October 26, 2004  vol. 101 no. 43</sub>]]
** T1
***5- CTCCCCG CGAAAAG AGTGTAC AGCGAAC GCCACG GTAAGCC TTCTTTC GCTTACC-3
** T2
***5- CTCCCCG CGAAAAG AGTGTAC AGCGAAC GCCACG GTAAGCC TTCTTTC GCTTACC-3
** T3
***5- CTCCCCG CGAAAAG AGTGTAC AGCGAAC GCCACG GTAAGCC TTCTTTC GCTTACC-3
** T4
***5- CTCCCCG CGAAAAG AGTGTAC AGCGAAC GCCACG GTAAGCC TTCTTTC GCTTACC-3
** Trigger
***5-CAGCGAACCTTTTCG AAAGCG GGGAG TTGCGTCCGTAT-3


* Version 2
= DNAzyme =
** T-1:
*** ACGGCGAGGAGGGTTAACCCAGACGCCGAATCCTAGACTCAAAGTAGTCTAGGATTCGGC<br>GTCTGAAAGGTAAGCCGTAGCTTCTTCTAATACGGCTTACC
** T-2:
*** CCATTCGGCATAATCTTCTTCGATGCCGAATGGTTTAGTCTAGGATTCGGCGTCTGGACTCTC<br>AGACGCCGAATCCTAGACTACTTTGGACAGGAGCGGC
** T-3:
*** ACGGCTTACCTTTAGAGTCCAGACGCCGAATCCTAGACTGGACTTAGTCTAGGATTCGGCGT<br>CTGCCCTCCTCGCCGTCAGCTTTCTCTAATACGGCGAGGA
** T-4:
*** AGGAGCGGCATAATCTCTTTCGATGCCGCTCCTGTCAGTCTAGGATTCGGCGTCTGAATTGG<br>CAGACGCCGAATCCTAGACTAAGTCCAAACCATTCGGCA
**Trigger:
***AGTCTAGGATTCGGCGTCTG GGTTAA


*Version 3
DNAzymes (also known as deoxyribozymes, DNA enzymes or catalytic DNA, are DNA molecules that have the ability to perform a chemical reaction, such as catalytic action. Since the description of the first DNAzyme for the cleavage of RNA in 1994, many more DNAzymes have been reported to catalyze many different types of chemical transformations, such as porphyrin metalation, DNA phosphorylation, RNA ligation, thymine-thymine dimer repair, carbon-carbon bond formation, and hydrolytic cleavage of DNA. DNA is chemically stable and can be conveniently produced by highly efficient automated DNA synthesis. Therefore, DNAzymes can be quite useful in research and applications in chemical biology, biotechnology, and medical areas.
**T1
***5-ATACGGACGCAACTCCCCGCTTTCGAAAAGGTTCGCTGAGTGTACAGCGAACCTTTTCGG<br>CCACGGTAAGCCGTATAGCTTCTTTCTAATACGGCTTACC-3
**T2
***5-GGTAAGCCGTATAGCTTCTTTCTAATACGGCTTACCGGTACCAGCGAACCTTTTCGAAAGC<br>G CGAAAAGGTTCGCTGTACACTCACGTTTGCGTCCGTAT-3
** T3
***5-ATACGGCTTACCGTGGCCGCTTTCGAAAAGGTTCGCTGAGTGTACAGCGAACCTTTTCGG<br>GGAG TTGCGTCCGTATAGCTCTTCTCTAATACGGACGCAA-3
** T4
***5-TTGCGTCCGTATAGCTCTTCTCTAATACGGACGCAAACGTGCAGCGAACCTTTTCGAAAGC<br>G CGAAAAGGTTCGCTGTACACTGTACCGGTAAGCCGTAT-3
** Trigger
***5-CAGCGAACCTTTTCG AAAGCG GGGAG TTGCGTCCGTAT-3


=The Walker =
[[Image:TJU-figure2.1.2.png|thumb|center|400px| '''Figure 2.1.2''' The sequence of 8-17 DNAzyme.The triangle represents the cutting site. (From Peracchi A <i>et al., J. Mol. Biol.</i>, 2005(352): 783–794.)]]


* Quadruped-walker
The Cu<sup>2+</sup> DNAzyme is also an ssDNA that contains a stem-loop of 8 base-pairing. The catalytic domain consists of a conservative sequence of six basepair. The two binding arms flanking the catalytic domain bind with the substrate, one of which forms a DNA triplex of the stem-loop with the substrate. Unlike 8-17, the substrate of Cu<sup>2+</sup> DNAzyme is deoxyribonucleotide. When the Cu<sup>2+</sup>concentration is <1μM, DNAzyme is still activated. When other ions’ concentration is enormously bigger than Cu<sup>2+</sup>, the DNAzyme still didn’t recover its full activity, which shows its great selectivity of Cu<sup>2+</sup>.
** WALKER1
***5-ATGGACTACGCTACCATTTTTTTTTTTTTTTTTTTTGGTAAGCCTGGGCCTCTTTCTTTTTAAG<br>AAAGAAC-3
** WALKER2
***5-TATAGCACGGTAAACTTAGTCCATTTTTTTTTTTTTTTTTTTTTGGTAAGCCTGGGCCTCTTTC<br>TTTTTAAGAAAGAAC-3
** WALKER3
***5-TGGTAGCGAGTTTACCTTACGGAGTTTTTTTTTTTTTTTTTTTTTTGCGTCCTGGGCCTCTCTT<br>CTTTTAGAAGAGAAC-3
** WALKER4
***5-CTCCGTAACTCGTATATTTTTTTTTTTTTTTTTTTTTTGCGTCCTGGGCCTCTCTTCTTTTAGA<br>AGAGAAC-3
** Two-foot-walker


* W2KER1
=DNA-walker =
**5-TTTCACATGCAATCGAACGTTTTTTTTTTTTGGTAAGCCTGGGCCTCTTTCTTTTTAAGAAAGA<br>AC-3
* W2KER2
**5-AACGTTCGATTGCATGTGAAATTTTTTTTTTTTGCGTCCTGGGCCTCTCTTCTTTTAGAAGAGA<br>AC-3


= Agarose electrophoresis =
DNA walkers are a class of nucleic acid nanomachines that exhibit directional motion along a linear track. A large number of schemes have been demonstrated. One strategy is to control the motion of the walker along the track using control strands that need to be manually added in sequence. Another approach is to make use of restriction enzymes or deoxyribozymes to cleave the strands and cause the walker to move forward, which has the advantage of running autonomously, and we choose this kind of walker this year. A later system could walk upon a two-dimensional surface rather than a linear track, and demonstrated the ability to selectively pick up and move molecular cargo.[55] Additionally, a linear walker has been demonstrated that performs DNA-templated synthesis as the walker advances along the track, allowing autonomous multistep chemical synthesis directed by the walker.


The mechanism of the DNA walker with a DNAzyme should be like this.


*0.2g agarose mixed with 20ml 1X TAE, heat to boil
[[Image:TJU-figure2.1.3.png|thumb|center|400px| '''Figure2.1.3''' The mechanism of the walker with DNAzyme.]]
*When temperature drops, add 0.1μL golden view, cooling for 30min
*Adding 1X TAE into
*Mix 2μL 6X loading buffer with 5μL DNA sample, add 7μL of the sample into the well
*Start the electrophoresis at 150-250V, for 10-30min, depending on the blue band of loading buffer
* Stop the electrophoresis, put the gel into UV gel imager, expose for 80ms


=PAGE=
=Origami=
DNA origami is the nanoscale folding of DNA to create arbitrary two and three dimensional shapes at the nanoscale. The specificity of the interactions between complementary base pairs make DNA a useful construction material, through design of its base sequences.


Developed by Paul Rothemund at the California Institute of Technology, the process involves the folding of a long single strand of viral DNA aided by multiple smaller "staple" strands. These shorter strands bind the longer in various places, resulting in various shapes, including a smiley face and a coarse map of China and the Americas, along with many three-dimensional structures such as cubes.


Nucleic acid electrophoresis utilizes the same concepts as protein electrophoresis. The denaturant and visualization techniques differ. All nucleic acids are negatively charged. It is not neccessary or desirable to use a charged denaturant such as SDS. Urea is used to denature the DNA or RNA within the gel. Visualization is usually achieved via staining with ethidium bromide. Ethidium bromide intercalates between base pairs and fluoresces. The fluoresence of free (non-bound) ethidium bromide is quenched. Note that ethidium bromide is a carcinogen and that UV light can damage skin and eyes. Gloves and protective eyeware should be used. The UV box has special anti-UV plastic and a safety interlock. Do not try to defeat the interlock.
To produce a desired shape, images are drawn with a raster fill of a single long DNA molecule. This design is then fed into a computer program that calculates the placement of individual staple strands. Each staple binds to a specific region of the DNA template, and thus due to Watson-Crick base pairing, the necessary sequences of all staple strands are known and displayed. The DNA is mixed, then heated and cooled. As the DNA cools, the various staples pull the long strand into the desired shape. Designs are directly observable via several methods, including atomic force microscopy, or fluorescence microscopy when DNA is coupled to fluorescent materials.


==Materials==
This year, we used the design from 2012 Harvard BIOMOD team to build the origami. What’s different is that we load ssDNAs on the staple strand. The ssDNAs can serve as the substrate of logic gate of 8-17 and Cu2+ DNAzyme, thus the release can be controlled by it. This new origami can serve as a miRNA delivery system based on ion detection.


Glass plates (10 x 20 cm), spacers, comb, and clamps
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Power supply
 
Nucleic Acid samples
 
6.66 mL 30% Acrylamide
 
3.34 mL 5X TBE
 
8.41 g Urea
 
The TA will have prepared the solution above before lab starts. The solution must be heated to dissolve the urea. However warm gel solutions polymerize quickly with unpredictable results, so the solution must be allowed to cool before use.
 
140 microliters 10% Ammonium Persulfate
 
7 microliters TEMED
 
Loading buffer- 2X Dye- 0.25% Bromophenol Blue, 0.25% Xylene Cyanol
 
Gel Red
 
1X TBE for Running Buffer
 
==Methods==
 
Very similar to SDS PAGE gel. You should be able to assemble gel by yourself. Have the TA inspect gel prior to use.
 
Add 8 microliters of the Loading Buffer to 10 microliters of your nucleic acid sample
 
Load onto gel, and separate your nucleic acid fragments by electrophoresis at around 30 Amps, until the dye front approaches the bottom of gel. Do NOT let dye front run off gel.
 
Stain with Gel Red for 20 minutes. Visualize on the UV box. Caution: Wear safety glasses around UV light.
 
Photograph the gel. Do not dispose of the gel until you have ensured that you have a reasonable photograph.
 
=PCR Annealing Procedure (for Logic Gate)=----
 
*90℃ for 5 min;
*90℃ for 1min;-1℃ per cycle
*Go to 2 for 86 times
*4℃ forever
 
=Recipe=
 
*DNAzyme Buffer
1M NaCl, 30mM HEPES, pH=7.0
*TBE
10x TBE (1 liter): Dissolve 108 g Tris and 55 g Boric acid in 900 ml distilled water. Add 40 ml 0.5 M Na2EDTA (pH 8.0) (alternatively use 9.3 g Na2EDTA) Adjust volume to 1 Liter. Store at room temperature. Note: 10x TBE may take some time to dissolve, even with fast stirring TBE can be diluted to 1X prior to use in electrophoresis, 0.5x is acceptable as well.
 
*TAE
TAE buffer is commonly prepared as a 50X stock solution for laboratory use. A 50X stock solution can be prepared by dissolving 242g Tris base in water, adding 57.1mL glacial acetic acid, and 100mL of 500mM EDTA (pH 8.0) solution, and bringing the final volume up to 1 liter. This stock solution can be diluted 50:1 with water to make a 1X working solution. This 1X solution will contain 40mM Tris, 20mM acetic acid, and 1mM EDTA.
 
*Urea Loading Buffer
8 M urea, 50 mM EDTA, 0.05% xylene cyanol, and 50 mM Tris acetate, pH 8.2.
 
 
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<br />
<br />
E-mail:austinamens@gmail.com | Address:Building No.20, No.92 Weijin Road, Tianjin, China | Zip-code:300072
E-mail:austinamens@gmail.com | Address:Building No.20, No.92 Weijin Road, Tianjin, China | Zip-code:300072
<br />Copyright 2013 © Tianjin University iGEM Team</p>
<br />Copyright 2013 © Tianjin University Biomod Team</p>
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   <li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin" onmouseover="mopen('m1')" onmouseout="mclosetime()" style="padding:15px 0px;">Home</a>

</li> <li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Team" onmouseover="mopen('m2')" onmouseout="mclosetime()" style="padding:15px 0px;">Team</a>

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</div> </li> <li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs" onmouseover="mopen('m3')" onmouseout="mclosetime()" style="padding:15px 0px;">Designs</a>

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               <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs">Background</a>

<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/Self-assemblyTrack">Self-assembly Track</a> <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/DeliveryDevice">The Delivery Device</a> </div> </li> <li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results" onmouseover="mopen('m4')" onmouseout="mclosetime()" style="height:35px;padding:8px 0px 7px 0px;">Experiments &amp; Results</a>

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<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results">Polymerizing</a> <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results/Cleavage">Cleavage</a> <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Experiments &amp; Results/DeliveryDevice">Delivery Device</a>

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   <li><a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Protocol" onmouseover="mopen('m5')" onmouseout="mclosetime()" style="padding:15px 0px;">Protocol</a>

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<a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs" style="font-size:14px;"><b>Background</b></a> <p>HCR、DNAzyme、DNA-walker、Origami</p> <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/Self-assemblyTrack" style="font-size:14px;"><b>Self-assembly Track</b></a> <p>T1-T4、The polymerization reaction</p> <a href="http://openwetware.org/wiki/Biomod/2013/Tianjin/Designs/DeliveryDevice"><b>The Delivery device</b></a> <br/><br/> </div>

<div class="photo rotate_right" style="width:400px;align:center;padding:30px 30px 30px 60px;"> <span style="color:#000;font-size:22px;"><b>Background</b></span> <img src="http://openwetware.org/images/c/cd/TJU-figure-2.jpg" alt="" width="360px" height="278px"/> </div>


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document.getElementById("toc").className="tocbefore"; } else{ document.getElementById("toc").className="tocafter"; } } window.onscroll=getScrollTop; </script> <div style="width:280px;float:left;"> <table id="toc" class="toc" summary="Contents" style="font-family:Calibri;float:right;font-size:120%;"><tbody><tr><td><div id="toctitle"><h2>Contents</h2></div> <ul> <li class="toclevel-1"><a href="#HCR"><span class="tocnumber">1</span> <span class="toctext">HCR</span></a></li> <li class="toclevel-1"><a href="#DNAzyme"><span class="tocnumber">2</span> <span class="toctext">DNAzyme</span></a></li> <li class="toclevel-1"><a href="#DNA-walker"><span class="tocnumber">3</span> <span class="toctext">DNA-walker</span></a></li> <li class="toclevel-1"><a href="#Origami"><span class="tocnumber">4</span> <span class="toctext">Origami</span></a> </ul> </td></tr></tbody></table> </div>


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HCR

Stable DNA monomers assemble only upon exposure to atarget DNA fragment. In the simplest version of this process, two stable species of DNA hairpins coexist in solution until the introduction of initiator strands triggers a cascade of hybridization events that yields nicked double helices analogous to alternating copolymers. The average molecular weight of the HCR products varies inversely with initiator concentration. Amplification of more diverse recognition events can be achieved by coupling HCR to aptamer triggers. This functionality allows DNA to act as an amplifying transducer for biosensing applications.

Figure2.1.1 (a–c) Secondary structure schematic of HCR function. Letters marked with * are complementary to the corresponding unmarked letter.(a) Hairpins H1 and H2 are stable in the absence of initiator I. (b) I nucleates at the sticky end of H1 and undergoes an unbiased strand displacement interaction to open the hairpin. (c) The newly exposed sticky end of H1 nucleates at the sticky end of H2 and opens the hairpin to expose a sticky end on H2 that is identical in sequence to I. Hence, each copy of I can propagate a chain reaction of hybridization events between alternating H1 and H2 hairpins to form a nicked double-helix, amplifying the signal of initiator binding. Figure and introduction from Robert M. Dirks and Niles A. Pierce PNAS October 26, 2004 vol. 101 no. 43

DNAzyme

DNAzymes (also known as deoxyribozymes, DNA enzymes or catalytic DNA, are DNA molecules that have the ability to perform a chemical reaction, such as catalytic action. Since the description of the first DNAzyme for the cleavage of RNA in 1994, many more DNAzymes have been reported to catalyze many different types of chemical transformations, such as porphyrin metalation, DNA phosphorylation, RNA ligation, thymine-thymine dimer repair, carbon-carbon bond formation, and hydrolytic cleavage of DNA. DNA is chemically stable and can be conveniently produced by highly efficient automated DNA synthesis. Therefore, DNAzymes can be quite useful in research and applications in chemical biology, biotechnology, and medical areas.

Figure 2.1.2 The sequence of 8-17 DNAzyme.The triangle represents the cutting site. (From Peracchi A et al., J. Mol. Biol., 2005(352): 783–794.)

The Cu2+ DNAzyme is also an ssDNA that contains a stem-loop of 8 base-pairing. The catalytic domain consists of a conservative sequence of six basepair. The two binding arms flanking the catalytic domain bind with the substrate, one of which forms a DNA triplex of the stem-loop with the substrate. Unlike 8-17, the substrate of Cu2+ DNAzyme is deoxyribonucleotide. When the Cu2+concentration is <1μM, DNAzyme is still activated. When other ions’ concentration is enormously bigger than Cu2+, the DNAzyme still didn’t recover its full activity, which shows its great selectivity of Cu2+.

DNA-walker

DNA walkers are a class of nucleic acid nanomachines that exhibit directional motion along a linear track. A large number of schemes have been demonstrated. One strategy is to control the motion of the walker along the track using control strands that need to be manually added in sequence. Another approach is to make use of restriction enzymes or deoxyribozymes to cleave the strands and cause the walker to move forward, which has the advantage of running autonomously, and we choose this kind of walker this year. A later system could walk upon a two-dimensional surface rather than a linear track, and demonstrated the ability to selectively pick up and move molecular cargo.[55] Additionally, a linear walker has been demonstrated that performs DNA-templated synthesis as the walker advances along the track, allowing autonomous multistep chemical synthesis directed by the walker.

The mechanism of the DNA walker with a DNAzyme should be like this.

Figure2.1.3 The mechanism of the walker with DNAzyme.

Origami

DNA origami is the nanoscale folding of DNA to create arbitrary two and three dimensional shapes at the nanoscale. The specificity of the interactions between complementary base pairs make DNA a useful construction material, through design of its base sequences.

Developed by Paul Rothemund at the California Institute of Technology, the process involves the folding of a long single strand of viral DNA aided by multiple smaller "staple" strands. These shorter strands bind the longer in various places, resulting in various shapes, including a smiley face and a coarse map of China and the Americas, along with many three-dimensional structures such as cubes.

To produce a desired shape, images are drawn with a raster fill of a single long DNA molecule. This design is then fed into a computer program that calculates the placement of individual staple strands. Each staple binds to a specific region of the DNA template, and thus due to Watson-Crick base pairing, the necessary sequences of all staple strands are known and displayed. The DNA is mixed, then heated and cooled. As the DNA cools, the various staples pull the long strand into the desired shape. Designs are directly observable via several methods, including atomic force microscopy, or fluorescence microscopy when DNA is coupled to fluorescent materials.

This year, we used the design from 2012 Harvard BIOMOD team to build the origami. What’s different is that we load ssDNAs on the staple strand. The ssDNAs can serve as the substrate of logic gate of 8-17 and Cu2+ DNAzyme, thus the release can be controlled by it. This new origami can serve as a miRNA delivery system based on ion detection.

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<div class="link"> <p><br /> <br /> E-mail:austinamens@gmail.com | Address:Building No.20, No.92 Weijin Road, Tianjin, China | Zip-code:300072 <br />Copyright 2013 © Tianjin University Biomod Team</p> </div>


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