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| − | <style>.nav-global ul li a{ font-size:0.85rem; } @media screen and (max-width: 865px) { .nav-global ul li a{ font-size:0.85rem; } }</style>
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| − | <div class="breadcrumb"></div>
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| − | <!-- <div class="share">
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| − | <a class="addthis_button_compact" tabindex="-1"><i class="icon icon-share"></i><span>Share</span></a>
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| − | <span></span></div>
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| − | -->
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| − | <div id="feature" class="row collapse text-light feature-footer-active">
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| − | <div class="inner-wrap medium-24 columns">
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| − | <!--TYPO3SEARCH_begin-->
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| − | <div class="carousel-container hide-pager" id="c4811">
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| − | <div class="carousel " id="slideshow-4811" data-cycle-log="false" data-cycle-auto-height="calc" data-cycle-prev="#prev-4811" data-cycle-next="#next-4811" data-cycle-pager="#cycle-pager-4811" data-cycle-easing="easeInOutQuad" data-cycle-fx="scrollVertUp" data-cycle-center-horz="true" data-cycle-speed="1000" data-cycle-timeout="8000" data-cycle-paused="true" data-cycle-slides="> div.slide">
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| − | <link rel="stylesheet" href="/Team:NUDT_CHINA/CSS3?action=raw&ctype=text/css" />
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| − | <!-- Quick initialize with theme color from first slide -->
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| − | <section class="big-header bh__height-size-3 text-light bh__video">
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| − | <div class="bh__background bh__background-color " style="background-color: rgb(3,13,38);background-image:url(https://static.igem.org/mediawiki/2016/thumb/3/3f/NUDT_CHINA2016_FILES-img-banner-3.0.png/800px-NUDT_CHINA2016_FILES-img-banner-3.0.png);background-size:100%;background-repeat:no-repeat;"></div>
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| − | <div class="bh__content ">
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| − | <div class="bh__content-inner-wrap">
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| − | <!--h3 class="bh__subtitle"><a href="http#/explore/whats-here/exhibits/brick-by-brick/"><span class="line-wrap"><span class="line"><span class="bh__info">Exhibit / </span>Brick by Brick</span></span></a></h3-->
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| − | <h1 class="bh__title ">
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| − | <!--<a href="#">-->
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| − | <span class="line-wrap">
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| − | <span class="line">Cancer,</span></span>
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| − | <br>
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| − | <span style="line-height:75px;" class="line-wrap">
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| − | <span style="line-height:75px;" class="line">No More Hiding</span></span>
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| − | <!--</a>-->
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| − | </h1>
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| − | <h2 style="font-size:36px;" class="bh__title ">
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| − | <!--<a href="#">-->
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| − | <span style="line-height:41px;" class="line-wrap">
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| − | <span style="line-height:41px;" class="line">Development of A Novel</span></span>
| + | |
| − | <span style="line-height:41px;" class="line-wrap">
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| − | <span style="line-height:41px;" class="line">Blood-MicroRNA Handy Detection System with CRISPR</span></span>
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| − | <!--</a>-->
| + | |
| − | </h2>
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| − | </div>
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| − | </div>
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| − | </div>
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| − | </section>
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| − | <!-- end .feature-ce --></div>
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| − | <div class="cycle-pager" id="cycle-pager-4811"></div>
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| − | </div>
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| − | <!--TYPO3SEARCH_end--></div>
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| − | <!-- end #feature -->
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| − | <div style="margin-top:80px;" id="content-wrap-ink"><!-- adjust in add_JS -->
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| − | <div class="row footer-link" style="">
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| − | | + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − |
| + | |
| − | <!--标题-->
| + | |
| − | <h2>
| + | |
| − | <span><span style="color:#7f1015">Abstract</span></span><hr />
| + | |
| − | </h2>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">This
| + | |
| − | | + | |
| − | model was created to evaluate the effectiveness of initial design, and offered
| + | |
| − | | + | |
| − | guidelines about how the system can (or must) be improved. (You can go <a href="/Team:NUDT_CHINA/Design">DESIGN.page</a></li> to see more.) The core idea was to simulate the process of producing the
| + | |
| − | | + | |
| − | signal which can be detected, and drew a conclusion by obtaining the
| + | |
| − | | + | |
| − | relationship between the signal intensity and the concentration of miRNA.
| + | |
| − | | + | |
| − | </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h2>
| + | |
| − | <span><span style="color:#7f1015">Introduction </span></span><hr />
| + | |
| − | </h2>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">Mathematical models of two aspects of our project were created, a
| + | |
| − | | + | |
| − | RCA model and a signal detection model.
| + | |
| − | | + | |
| − | </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The RCA
| + | |
| − | | + | |
| − | model was based on the Michaelis-Menten equation. The relationship between the
| + | |
| − | | + | |
| − | concentration of miRNA and the number of stem-loop structures was obtained
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| − | | + | |
| − | through theoretical calculation, and our experimental results was used to
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| − | | + | |
| − | calculate the parameters we introduced previously.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | signal detection model was composed of a probability model and a kinetic
| + | |
| − | | + | |
| − | equation of enzymatic reaction, thus the relationship between the number of
| + | |
| − | | + | |
| − | stem-loop structures and the signal intensity under the premise of adding a
| + | |
| − | | + | |
| − | certain amount of fusion proteins of dCas9 and split-HRP fragments was obtained.
| + | |
| − | | + | |
| − | </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">By
| + | |
| − | | + | |
| − | integrating the two models, the impacts of the molecule number of proteins on
| + | |
| − | | + | |
| − | the signal to noise ratio was theoretically predicted and the trend of the
| + | |
| − | | + | |
| − | signal intensity with the change of the concentration of miRNA in our wet-lab
| + | |
| − | | + | |
| − | work was explained.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h2>
| + | |
| − | <span><span style="color:#7f1015"> Assumption and Justification
| + | |
| − | | + | |
| − | </span></span><hr />
| + | |
| − | </h2>
| + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015"> About model </span></span><hr />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">
| + | |
| − | | + | |
| − | Assuming that miRNA is not degraded throughout the reaction process.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">
| + | |
| − | | + | |
| − | Assuming that the two fusion proteins of dCas9 and split-HRP fragments have the
| + | |
| − | | + | |
| − | same ability to combine with the stem-loop structure, and only when two
| + | |
| − | | + | |
| − | different proteins next to each other, can they have the ability to catalyze
| + | |
| − | | + | |
| − | substrate and produce signal.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">
| + | |
| − | | + | |
| − | Assuming that the number of stem-loop structures in each RCA product is equal
| + | |
| − | | + | |
| − | under a certain reaction time. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">
| + | |
| − | Assuming that the enzymatic activity remains unchanged with time under the
| + | |
| − | | + | |
| − | premise of excessive amount of enzymes or a short-time reaction.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015"> About the data </span></span><hr
| + | |
| − | | + | |
| − | />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;"> Assuming that the data we obtain from wet-lab experiment are
| + | |
| − | | + | |
| − | reliable.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;"> Assuming that all the results are trustworthy in the process of
| + | |
| − | | + | |
| − | statistical processing and data calculation.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h2>
| + | |
| − | <span><span style="color:#7f1015">Model </span></span><hr />
| + | |
| − | </h2>
| + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015">Notations </span></span><hr />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--表格-->
| + | |
| − | <p>
| + | |
| − | <table class="MsoTableGrid" border="1" cellspacing="0" cellpadding="0" style="border:none;">
| + | |
| − | <tbody>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>Symbol </span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>Definition </span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>X</span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK7"></a><a name="OLE_LINK6"></a><span>The concentration of</span><span> miRNA (pM)</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>C<sub>1</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The concentration of initiated probe (Abbreviated
| + | |
| − | to iprobe)</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>k<sub>1</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK8"></a><span>A constant representing
| + | |
| − | the scale factor</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>K<sub>m </sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>One of the characteristic constants of
| + | |
| − | phi29 DNA polymerase</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>V<sub>max</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>One of the characteristic constants of phi29
| + | |
| − | DNA polymerase</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>k<sub>2</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>A constant representing the scale factor</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>V</span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The initial speed of RCA </span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK9"></a><i><span>n<sub>1</sub></span></i><i><sub><span></span></sub></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The moles of RCA product</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>n<sub>2</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK10"></a><span>The number of
| + | |
| − | stem-loop structures in each RCA product</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>n</span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The total amount of <a name="OLE_LINK23"></a><a name="OLE_LINK22"></a>stem-loop structures</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>N</span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK41"></a><a name="OLE_LINK16"></a><span>The molecule number of fusion
| + | |
| − | protein of dCas9 and split-HRP fragments</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>k<sub>3</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>A constant representing the scale factor</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>y<sub>1</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The fluorescence intensity of
| + | |
| − | DNA-dye-complex (RFU)</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>N<sub>1</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The molecule number of fusion protein of
| + | |
| − | dCas9 and split-HRP fragments in the solution</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>N<sub>2</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK45"></a><span>The molecule number
| + | |
| − | of fusion protein of dCas9 and split-HRP fragments binding with stem-loop
| + | |
| − | structure</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>k<sub>4</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>A constant representing the scale factor</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span>k<sub>5</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>A constant representing the scale factor</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span style="font-family:宋体;">ρ</span><span></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>Signal to noise ratio(Abbreviated to
| + | |
| − | SNR)</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span style="font-family:宋体;">I</span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The molecule number of formed intact HRP
| + | |
| − | proteins </span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span style="font-family:宋体;">I<sub>1</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The molecule number of formed intact HRP
| + | |
| − | proteins in the solution</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span style="font-family:宋体;">I<sub>2</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <a name="OLE_LINK43"></a><span>The molecule number
| + | |
| − | of formed intact HRP proteins through binding with stem-loop structure</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span style="font-family:宋体;">t</span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>Reaction time</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td width="66" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <i><span style="font-family:宋体;">y<sub>2</sub></span></i>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | <td width="487" valign="top" style="border:solid windowtext 1.0pt;">
| + | |
| − | <p class="MsoNormal">
| + | |
| − | <span>The signal intensity (OD<sub>450</sub>)</span>
| + | |
| − | </p>
| + | |
| − | </td>
| + | |
| − | </tr>
| + | |
| − | </tbody>
| + | |
| − | </table>
| + | |
| − | </p>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015"> The RCA model </span></span><hr
| + | |
| − | | + | |
| − | />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">Firstly, we assume that there is a linear relationship between the
| + | |
| − | | + | |
| − | concentration of the initiated probes and miRNA as the combination reaction
| + | |
| − | | + | |
| − | between miRNA and probe occurs spontaneously<sup>1 </sup> . This can be written
| + | |
| − | | + | |
| − | as:
| + | |
| − | | + | |
| − | </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model1.jpg|70px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">Next,
| + | |
| − | | + | |
| − | RCA is an enzymatic reaction, the reaction equation is written as: iprobe+dNTP→
| + | |
| − | | + | |
| − | iprobe-dN+PPi, under the premise of excessive amount of enzymes and dNTPs, the
| + | |
| − | | + | |
| − | relationship between the concentration of iprobe and the initial speed of RCA
| + | |
| − | | + | |
| − | can be described by the Michaelis-Menten equation. This can be written as：
| + | |
| − | | + | |
| − | </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model2.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">It is
| + | |
| − | | + | |
| − | obvious that there is a linear relationship between the molecule numbers of
| + | |
| − | | + | |
| − | iprobe-dN (n<sub>1 </sub>, reflects the molecule numbers of RCA product) and the initial
| + | |
| − | | + | |
| − | speed of RCA reaction. Notably, under the premise of excessive amount of
| + | |
| − | | + | |
| − | enzymes, the extending of each RCA production is related to the reaction time,
| + | |
| − | | + | |
| − | not the concentration of the iprobe. It is a linear relationship when we assume
| + | |
| − | | + | |
| − | that the enzymatic activity remains unchanged with time, thus, the number of
| + | |
| − | | + | |
| − | stem-loop structures (n<sub>2 </sub>) in each RCA product is stable under the premise of a
| + | |
| − | | + | |
| − | certain reaction time. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig1.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--图注-->
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure 1. Schematic diagram</span></b>
| + | |
| − | </p>
| + | |
| − | | + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">This
| + | |
| − | | + | |
| − | can be written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model3.jpg|70px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model4.jpg|70px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">And
| + | |
| − | | + | |
| − | then, SYBR Green binds to DNA. The resulting DNA-dye-complex absorbs blue light
| + | |
| − | | + | |
| − | (λmax = 497nm) and emits green light (λmax = 520nm). Thus, there is a linear
| + | |
| − | | + | |
| − | relationship between the total amount of stem-loop structures and the
| + | |
| − | | + | |
| − | fluorescence intensity of DNA-dye-complex. This can be written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model5.jpg|70px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">In
| + | |
| − | | + | |
| − | summary, the relationship between the concentration of miRNA and the
| + | |
| − | | + | |
| − | fluorescence intensity of DNA-dye-complex can be written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model6.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015"> The signal detection model
| + | |
| − | | + | |
| − | </span></span><hr />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Firstly, it is obvious that there is a linear relationship between
| + | |
| − | | + | |
| − | the molecule number of formed intact HRP proteins in the solution (consider as
| + | |
| − | | + | |
| − | NOISE) and the molecule number of fusion protein s in the solution. This can be
| + | |
| − | | + | |
| − | written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model7.jpg|70px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">Next,
| + | |
| − | | + | |
| − | we built a probability model to find the expression of the molecule number of
| + | |
| − | | + | |
| − | formed intact HRP proteins (fused with dCas9) through binding with stem-loop
| + | |
| − | | + | |
| − | structure (consider as SIGNAL). It is related to the total amount of stem-loop
| + | |
| − | | + | |
| − | structures and the molecule number of DNA-bound fusion proteins. We obtain the
| + | |
| − | | + | |
| − | result by Monte Carlo method. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--图和图注-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig2.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure2. Frequency Distribution</span></b>
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:16px;">Plot of
| + | |
| − | | + | |
| − | frequency distribution of the molecule number of formed intact HRP proteins
| + | |
| − | | + | |
| − | through binding with stem-loop structure. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--图和图注-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig3.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure 3. The result of the probability model</span></b>
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:16px;">Plot of
| + | |
| − | | + | |
| − | the expect value of the molecule number of formed intact proteins against the
| + | |
| − | | + | |
| − | total amount of stem-loop structures, carried out at 0.1, 0.3, 0.5, 0.7, 0.9 of
| + | |
| − | | + | |
| − | N2 to n ratio. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | result can be written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model8.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">As for
| + | |
| − | | + | |
| − | N2, which refers to the molecule number of fusion proteins binding with stem-
| + | |
| − | | + | |
| − | loop structure. The equation is formulated based on the limiting case. The
| + | |
| − | | + | |
| − | equation can be written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model9.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">And
| + | |
| − | | + | |
| − | then, we can obtain the expressions for ρ and I separately.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model10.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model11.jpg|70px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Notably, under the premise of excessive substrates, it is a linear
| + | |
| − | | + | |
| − | relationship between the reaction rate and the concentration of enzymes when we
| + | |
| − | | + | |
| − | assume that the enzymatic activity remain unchanged with time, thus, we can
| + | |
| − | | + | |
| − | obtain the expression of the signal intensity at tth time-step. This is written
| + | |
| − | | + | |
| − | as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model12.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">In
| + | |
| − | | + | |
| − | summary, the relationship between the number of stem-loop structures and the
| + | |
| − | | + | |
| − | signal to noise ratio under the premise of adding a certain amount of fusion
| + | |
| − | | + | |
| − | proteins is written as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model13.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | relationship between the number of Stem-loop structures and the signal intensity
| + | |
| − | | + | |
| − | under the premise of adding a certain amount of fusion proteins is written
| + | |
| − | | + | |
| − | as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model14.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015"> The calculation of the constants
| + | |
| − | | + | |
| − | </span></span><hr />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">To
| + | |
| − | | + | |
| − | simplify the equation (1), the constants could be integrated. As can be seen
| + | |
| − | | + | |
| − | from the above table, k1, k2, k3 are constants representing the scale factor, Km
| + | |
| − | | + | |
| − | and Vmax are characteristic constants of phi 29 DNA polymerase, and n2, which
| + | |
| − | | + | |
| − | refers to the number of stem-loop structures in each RCA product, is stable
| + | |
| − | | + | |
| − | under the premise of a certain reaction time. Thus, define two constants, mark
| + | |
| − | | + | |
| − | as a and b, and then the equation can be simplified as:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--插入图片-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--model15.jpg|105px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">We used
| + | |
| − | | + | |
| − | this equation to fit the data points obtained through experiments. (Figure …in
| + | |
| − | | + | |
| − | the ….page) The fitting curve is shown below.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--图和图注-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig4.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure 4. Fitting curve</span></b>
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:16px;">Fitting
| + | |
| − | | + | |
| − | curve of fluorescence intensity of DNA-dye-complex (RFU) against the
| + | |
| − | | + | |
| − | concentrations of miRNA (Pm). </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">With
| + | |
| − | | + | |
| − | regard to the equation (2), we set the parameters based on the experiments as
| + | |
| − | | + | |
| − | follows:
| + | |
| − | k3 =8.35*10-16 (n=y1/k3); k4 =0.038; k5t=5.9*10-12.
| + | |
| − | </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h3>
| + | |
| − | <span><span style="color:#7f1015"> Results and Analysis
| + | |
| − | | + | |
| − | </span></span><hr />
| + | |
| − | </h3>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">By
| + | |
| − | | + | |
| − | integrating the two models, we can obtain the relationships between the signal
| + | |
| − | | + | |
| − | to noise ratio, the signal intensity respectively and the concentration of miRNA
| + | |
| − | | + | |
| − | under different additional amount of fusion proteins.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | results are shown below.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--图和图注-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig5.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure 5. The signal intensity (OD450)</span></b>
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:16px;">Three-
| + | |
| − | | + | |
| − | dimensional map of signal intensity (OD450) against miRNA concentration(pM) and
| + | |
| − | | + | |
| − | additional amount of fusion proteins. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">There
| + | |
| − | | + | |
| − | exists a monotonous relation between the concentration of miRNA and the signal
| + | |
| − | | + | |
| − | intensity when the value of the molecule number of fusion proteins is relatively
| + | |
| − | | + | |
| − | large. While the relationship does not hold when the value of the molecule
| + | |
| − | | + | |
| − | number of fusion proteins is relatively small. And the signal intensity
| + | |
| − | | + | |
| − | increases as the value of the molecule number of fusion proteins
| + | |
| − | | + | |
| − | increases.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | trend of the signal intensity with the change of the concentration of miRNA is
| + | |
| − | | + | |
| − | consistent with that we obtained in our wet-lab work.(…….. in the result
| + | |
| − | | + | |
| − | page)</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--图和图注-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig6.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure 6. The result of signal to noise ratio</span></b>
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:16px;">Three-
| + | |
| − | | + | |
| − | dimensional map of signal to noise ratio against miRNA concentration (pM) and
| + | |
| − | | + | |
| − | additional amount of fusion proteins.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | signal-to-noise ratio decreases as the value of the molecule number of fusion
| + | |
| − | | + | |
| − | proteins increases. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Furthermore, we consider that it is conducive to signal detection
| + | |
| − | | + | |
| − | when the values of signal intensity and signal to noise ratio are both greater
| + | |
| − | | + | |
| − | than two. We obtain the ideal region through calculation, which is shown as
| + | |
| − | | + | |
| − | below. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--图和图注-->
| + | |
| − | </br>
| + | |
| − | </html>
| + | |
| − | [[File:T--NUDT_CHINA--modelfig7.jpg|350px|center]]
| + | |
| − | <html>
| + | |
| − | </br>
| + | |
| − | <p>
| + | |
| − | <b><span style="line-height:2;font-family:Perpetua;font-
| + | |
| − | | + | |
| − | size:18px;">Figure 7. Ideal region</span></b>
| + | |
| − | </p>
| + | |
| − | <p>
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:16px;">Region
| + | |
| − | | + | |
| − | of qualified logarithm of concentrations of miRNA (pM) and logarithm of the
| + | |
| − | | + | |
| − | additional amount of fusion proteins.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">When
| + | |
| − | | + | |
| − | the value of N is set to 1013.3, the range of the value of x contained in the
| + | |
| − | | + | |
| − | ideal region is the largest, which means when the value of the molecule number
| + | |
| − | | + | |
| − | of fusion proteins is 1013.3, the concentration range of miRNA contained in the
| + | |
| − | | + | |
| − | ideal region is the largest. </span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--标题-->
| + | |
| − | <h2>
| + | |
| − | <span><span style="color:#7f1015">Conclusion </span></span><hr />
| + | |
| − | </h2>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">Based
| + | |
| − | | + | |
| − | on our simulation, we came to the conclusion that:</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | calculated trend of the signal intensity with the change of the concentration of
| + | |
| − | | + | |
| − | miRNA from our model is consistent with the trend we obtained in our laboratory
| + | |
| − | | + | |
| − | work.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
| − | | + | |
| − | <!--正文-->
| + | |
| − | <p style="text-indent:22pt;">
| + | |
| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
| + | |
| − | | + | |
| − | value of the molecule number of fusion protein of dCas9 and split-HRP fragments
| + | |
| − | | + | |
| − | not only affects the signal-to-noise ratio, but also the signal intensity. So we
| + | |
| − | | + | |
| − | need to weigh its impact on both to select the optimal solution.</span>
| + | |
| − | </p>
| + | |
| − | </br>
| + | |
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| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">The
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| − | concentration range of miRNA contained in the ideal region is the largest when
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| − | we set the value of the molecule number of fusion proteins to be 1013.3, we
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| − | optimize the value of the addition amount of fusion proteins in our wet-lab work
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| − | based on it.</span>
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| − | <span><span style="color:#7f1015">Reference </span></span><hr />
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| − | <!--正文-->
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| − | <p style="text-indent:22pt;">
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| − | <span style="line-height:2;font-family:Perpetua;font-size:18px;">1
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| − | Deng, R. et al. Toehold-initiated rolling circle amplification for visualizing
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| − | individual microRNAs in situ in single cells. Angew Chem Int Ed Engl 53, 2389-
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| − | 2393, doi:10.1002/anie.201309388 (2014).</span>
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