Biomod/2012/TeamSendai/Design: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
No edit summary
Line 9: Line 9:
[[Image: Designtopcellgate.png |left|340px]]
[[Image: Designtopcellgate.png |left|340px]]
Our project "Cell-gate" is left.
Our project "Cell-gate" is left.
Our project is divided major three parts, Gate, Porter, and Membrane.
Our project is divided into major three parts, Gate, Porter, and Membrane.
Gate is Cell-gate itself.
Gate is Cell-gate itself.
　Porter is function to transport the target inside and outside cell membrane.
　Porter is function to transport the target inside and outside cell membrane.

Revision as of 03:59, 27 October 2012

<html> <head> 

   <title>Team Sendai Top</title>
   <script type="text/javascript" src="http://code.jquery.com/jquery-latest.min.js"></script>
   <script type="text/javascript">
       $(function() {
         if ($.browser.msie && $.browser.version.substr(0,1)<7)
         {

$('li').has('ul').mouseover(function(){ $(this).children('ul').show(); }).mouseout(function(){ $(this).children('ul').hide(); }) 

         }
       });        
   </script>

<style>

body{ background-image:url(http://openwetware.org/images/7/78/Office.png); /*壁色と壁紙設定*/ background-repeat:repeat;/*繰り返さない場合はno-repeatに変更*/ font:"メイリオ", "ＭＳ Ｐゴシック", Osaka, "ヒラギノ角ゴ Pro W3"; color: #333333; margin:0px; padding:0px; }

  1. Container

{ Background-image: url(http://openwetware.org/images/6/62/Wall4.png); width: 960px; margin: auto; }

/* Main menu */

  1. menu

{ width: 100%; margin: 0; padding: 10px 0 0 0; list-style: none; background: #111; background: -moz-linear-gradient(#444, #111); 

   background: -webkit-gradient(linear,left bottom,left top,color-stop(0, #111),color-stop(1, #444));

background: -webkit-linear-gradient(#444, #111); background: -o-linear-gradient(#444, #111); background: -ms-linear-gradient(#444, #111); background: linear-gradient(#444, #111); -moz-border-radius: 50px; border-radius: 50px; -moz-box-shadow: 0 2px 1px #9c9c9c; -webkit-box-shadow: 0 2px 1px #9c9c9c; box-shadow: 0 2px 1px #9c9c9c; }

  1. menu li

{ float: left; padding: 0 0 10px 0; position: relative; line-height: 0; }

  1. menu a

{ float: left; height: 25px; padding: 0 25px; color: #999; text-transform: uppercase; font: bold 12px/25px Arial, Helvetica; text-decoration: none; text-shadow: 0 1px 0 #000; }

  1. menu li:hover > a

{ color: #fafafa; }

  • html #menu li a:hover /* IE6 */

{ color: #fafafa; }

  1. menu li:hover > ul

{ display: block; }

/* Sub-menu */

  1. menu ul

{ 

   list-style: none;
   margin: 0;
   padding: 0;    
   display: none;
   position: absolute;
   top: 35px;
   left: 0;
   z-index: 99999;    
   background: #444;
   background: -moz-linear-gradient(#444, #111);
   background: -webkit-gradient(linear,left bottom,left top,color-stop(0, #111),color-stop(1, #444));
   background: -webkit-linear-gradient(#444, #111);    
   background: -o-linear-gradient(#444, #111);	
   background: -ms-linear-gradient(#444, #111);	
   background: linear-gradient(#444, #111);
   -moz-box-shadow: 0 0 2px rgba(255,255,255,.5);
   -webkit-box-shadow: 0 0 2px rgba(255,255,255,.5);
   box-shadow: 0 0 2px rgba(255,255,255,.5);	
   -moz-border-radius: 5px;
   border-radius: 5px;

}

  1. menu ul ul

{ 

 top: 0;
 left: 150px;

}

  1. menu ul li

{ 

   float: none;
   margin: 0;
   padding: 0;
   display: block;  
   -moz-box-shadow: 0 1px 0 #111111, 0 2px 0 #777777;
   -webkit-box-shadow: 0 1px 0 #111111, 0 2px 0 #777777;
   box-shadow: 0 1px 0 #111111, 0 2px 0 #777777;

}

  1. menu ul li:last-child

{ 

   -moz-box-shadow: none;
   -webkit-box-shadow: none;
   box-shadow: none;

}

  1. menu ul a

{ 

   padding: 10px;

height: 10px; width: 130px; height: auto; 

   line-height: 1;
   display: block;
   white-space: nowrap;
   float: none;

text-transform: none; }

  • html #menu ul a /* IE6 */

{ height: 10px; }

  • first-child+html #menu ul a /* IE7 */

{ height: 10px; }

  1. menu ul a:hover

{ 

   background: #0186ba;

background: -moz-linear-gradient(#04acec, #0186ba); background: -webkit-gradient(linear, left top, left bottom, from(#04acec), to(#0186ba)); background: -webkit-linear-gradient(#04acec, #0186ba); background: -o-linear-gradient(#04acec, #0186ba); background: -ms-linear-gradient(#04acec, #0186ba); background: linear-gradient(#04acec, #0186ba); }

  1. menu ul li:first-child > a

{ 

   -moz-border-radius: 5px 5px 0 0;
   border-radius: 5px 5px 0 0;

}

  1. menu ul li:first-child > a:after

{ 

   content: '';
   position: absolute;
   left: 30px;
   top: -8px;
   width: 0;
   height: 0;
   border-left: 5px solid transparent;
   border-right: 5px solid transparent;
   border-bottom: 8px solid #444;

}

  1. menu ul ul li:first-child a:after

{ 

   left: -8px;
   top: 12px;
   width: 0;
   height: 0;
   border-left: 0;	
   border-bottom: 5px solid transparent;
   border-top: 5px solid transparent;
   border-right: 8px solid #444;

}

  1. menu ul li:first-child a:hover:after

{ 

   border-bottom-color: #04acec;

}

  1. menu ul ul li:first-child a:hover:after

{ 

   border-right-color: #04acec; 
   border-bottom-color: transparent;

}


  1. menu ul li:last-child > a

{ 

   -moz-border-radius: 0 0 5px 5px;
   border-radius: 0 0 5px 5px;

}

/* Clear floated elements */

  1. menu:after

{ visibility: hidden; display: block; font-size: 0; content: " "; clear: both; height: 0; }

  • html #menu { zoom: 1; } /* IE6 */
    first-child+html #menu { zoom: 1; } /* IE7 */

/* 目次 */ div#mokuji {width: 960px; margin-left: auto; margin-right: auto; background-color: #f5f5dc}

div#mokuji h2 { background-color: # f5f5dc; font-size: 1.50em; color: #000000; line-height: 45px; padding-left: 12px; margin-bottom: 0}

div#mokuji h3 {border-bottom:solid 3px #66aa66; font-size: 1.50em; line-height: 22px; padding-left: 12px; margin-top: 30px; margin-bottom: 0; clear: both}


ol#mokuji {font-size: 1.00em; margin-left: 0; padding-left: 0}


/*文字の設定*/ h1{ font-family: 'Times New Roman', 'Times', serif; font-size: 2.00em; letter-spacing: 2px; padding-left:10px; margin: 0px; }


h2{ font-family: 'Times New Roman', 'Times', serif; 

     	font-size:1.8em;

border-left: 10px solid #000066; 

         	border-bottom:solid 1px #000099;/*文字の下に線を入れる設定*/
         	width:900px;

padding-left: 5px; color:#333333; margin-top: 15px; margin-bottom: 5px; }

h3{ font-family: 'Times New Roman', 'Times', serif; 

     font-size:1.60em;
       color:# 333333;

}

p{ 

   	font-family: 'Times New Roman', 'Times', serif  	font-size:1.40em;/*全体の文字サイズ*/

line-height:150%;/*全体で使う、文字と文字の行間*/ 

         	margin-left:5px;

}

p img{ 

    	float:left;
        	margin-top:5px;  /*写真の左にスペースを空ける*/

margin-left:5px; /*写真の左にスペースを空ける*/ margin-right:10px; /:写真と文字の間隔*/ }


h5{ 

     font-size:1.60em;

border: solid 1px #111111; 

       color:#ffffff;

background-color:#FFA500; line-height: 30px; padding-left:10px; margin-top: 10px; margin-bottom: 1px; }

h6{ 

     font-size:120%;

border: solid 1px #111111; 

       color:#ffffff;

background-color:#006400; line-height: 30px; padding-left:10px; margin-top: 10px; margin-bottom: 1px; }

  1. red{
     font-size:120%;

border: solid 1px #111111; 

       color:#ffffff;

background-color:#DC143C; line-height: 40px; padding-left:10px; margin-top: 10px; margin-bottom: 1px; }

  1. blue{
     font-size:120%;

border: solid 1px #111111; 

       color:#ffffff;

background-color:#191970; line-height: 40px; padding-left:10px; margin-top: 10px; margin-bottom: 1px; }

  1. pop {

width: 50%; margin: 0px; float: left; }

  1. pop h2 {

width: 90%; margin: 0px; float: left; }

  1. pop p {

width: 90%; margin: 0px; float: left; }


/*topに戻る*/

  1. ttop {position:fixed;bottom:140px;left:auto;margin:0 0 0 960px; /* マージン：上 右 下 左 */width:41px;height:390px;background:url(http://openwetware.org/images/e/ed/Cellgate.gif) no-repeat left bottom;}/* IE6以下用 */* html #ttop {margin:0 0 -390px 0;position:relative;bottom:490px; /* 上で設定した ttopの高さ390px＋下100px */left:960px;}#ttop:hover {background:url(http://openwetware.org/images/a/a7/029.png) no-repeat left bottom;/* 画像の高さによって適当に調整 */}a.page_top {display:block;width:41px;height:390px;}


</style> </head>


<body> <div id="Container"> <!-- Menu --> <ul id="menu"> <li><a href="http://openwetware.org/wiki/Biomod/2012/Tohoku/Team_Sendai ">Home</a></li> <li><a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Idea ">Project</a></li> <li><a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Design">Design</a> </li> <li><a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Simulation">Simulation</a> </li> <li> <a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Experiment ">Experiment</a> </li> <li> <a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Future Application">Future Application</a> </li> <li> <a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Diary">Diary</a> </li> <li> <a href=" http://openwetware.org/wiki/Biomod/2012/TeamSendai/Team ">Team</a> </li> <li> <a href=" http://openwetware.org/wiki/Biomod/2012/Tohoku/Team Sendai/header"></a> </li>

</ul>

<div id="ttop"> <a href="#top" class="page_top" onfocus="this.blur();" onclick="scrollTo(0,0); return false;" title="Top"></a></div>

</body> </div> </html>


Cell-gate

Our project "Cell-gate" is left. Our project is divided into major three parts, Gate, Porter, and Membrane. Gate is Cell-gate itself. 　Porter is function to transport the target inside and outside cell membrane. 　Membrane is liposome which is model of cell membrane. We divided our project into above three group and did experiment. On this page, we descript how we determined our robot design.

Design

Design of Gate

Size / Structure

The Gate has to connect inside and outside of the cell. So we decided to apply a hexagonal tube nanostructure made of DNA origami.

We refer "A logic-gated nanorobot for targeted transport of molecular payloads" (SM Douglas, I Bachelet, GM Church - Science Signalling, 2012) for the hexagonal tube structure of DNA origami.

Next, we made a simulation in order to examine the size of the structure. The size of the tube must be small enough not to pass freely through anything. However, it must be large enough to pass through the desired product. The gate which made of DNA origami has negative electric charge. So if the gate is too small, target can't enter the Gate. According to simulation, our Gate size determined 24*24*33nm. This size is suitable to transport the target.

DNA origami

We used caDNAno to design the hexagonal tube structure.



Potential Barrier

Our Gate is made of DNA, so it has negative electric charge. Single stranded DNA has negative electric charge, too. Here is a graph about potential energy of the tube.(藤原さんに頂いた下のグラフ載せる。) GATE size means the length of the Gate. If the potential energy is high, it is difficult for single stranded DNAs to enter the Gate. If the radius of the Gate is 1.5 times larger than now design, potential energy decreases and to enter the Gate is easier. So our Gate is well designed.


Design of Porter

Principle

When we consider making Cell Gate, there are two problems. One is how to pull the target oligonucleotide into Gate, and the other is how to pass the target. To solve these problems, we propose a nano structure of ssDNA and named it “Porter”. Porter stands in line in Gate, pulls the target, and transports it.

The Gate simulation shows that things by itself don’t come in Gate. So Porter has to pull and carry the target inside Gate.

We designed Porter having some loop structures when it hybridizes with the target. Porter has some complementary sequences to the target here and there. So after the target attaches to Porter at the end of Porter, Porter shrinks and pulls the target. Finally the target comes in Gate, otherwise unable to go through.

The inner Porter has more complementary sequences to the target and has higher bonding energy from the entrance of Gate. This design enables the target to move to the inner Porter because of the combination stability. In experiment, we apply the sequences below.

<DNA sequences>(実際の配列をお願いします) These are the sequences for electrophoresis, and only a part of the actual Porter sequence. These are the extracted region binding to the target. When planted in the gate, Porter has more spacer sequences of 10base at the root in addition to the sequences above, to reduce the Coulomb's force produced by the wall of Gate.


Simulation

We compared the ability to catch the target of Porter with that of toehold structure.

How to implement

Cell model

Cholesterol

We also have the aim of this GATE stab in the cell membrane, it can not sting in the cell membrane of normal hexagonal tube. However, We can create a tube with a different structure by exchanging some staple. We have designed a simple tube first,and I have to be attached anywhere in the structure to replace the staple. We can later add functionality to an existing structure using this method. We thought that to have an affinity for lipid membrane with the DNA that can be modified cholesterol on the side of the tube by this method. In addition, we have also designed DNA-like beard at the entrance of the tube. We expect the effect of electrostatically repel force with DNA which are not intended. Our tube is small,so we designed the tube to connect to each other and be long in order to easily confirmed using AFM. By replacing the staple, this structure is also removably.

Retrieved from "https://openwetware.org/mediawiki/index.php?title=Biomod/2012/TeamSendai/Design&oldid=645229"