Difference between revisions of "Team:SDU CHINA/Safety"

(Prototype team page)
 
Line 1: Line 1:
 
{{SDU_CHINA}}
 
{{SDU_CHINA}}
<html>
 
  
 +
<html lang="en">
 +
<head>
 +
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
 +
<style>
 +
.top{text-align:center}
 +
.p{text-indent:1.5em;word-wrap:break-word}
 +
.tu{text-align:center;font-size:1em}
 +
.all{margin:0 15% auto 15%;font-family: Segoe, "Segoe UI", "DejaVu Sans", "Trebuchet MS", Verdana, "sans-serif";letter-spacing:0.05em}
 +
.title{font-size:2em;text-align:center;font-family:Constantia, "Lucida Bright", "DejaVu Serif", Georgia, "serif"}
 +
.subtitle{text-align:center;font-size:1.2em;font-family: Baskerville, "Palatino Linotype", Palatino, "Century Schoolbook L", "Times New Roman", "serif"}
 +
.img{text-align:center;height:20em}
 +
.table{margin:0 auto}
 +
.STYLE1 {text-align: center; font-size: 1.2em; font-family: Baskerville, "Palatino Linotype", Palatino, "Century Schoolbook L", "Times New Roman", "serif"; font-weight: bold; }
 +
</style>
 +
</head>
  
<div class="column full_size">
+
<body>
 +
<br><br>
 +
<div class="top"><img src="https://static.igem.org/mediawiki/2017/d/d4/Safety.jpeg"></div>
 +
<br><br><br>
  
<h1> Safety </h1>
+
<div class="all">
<p>Please visit <a href="https://2017.igem.org/Safety">the main Safety page</a> to find this year's safety requirements & deadlines, and to learn about safe & responsible research in iGEM.</p>
+
  
<p>On this page of your wiki, you should write about how you are addressing any safety issues in your project. The wiki is a place where you can <strong>go beyond the questions on the safety forms</strong>, and write about whatever safety topics are most interesting in your project. (You do not need to copy your safety forms onto this wiki page.)</p>
+
<div>
 +
<div class="title" >
 +
  <strong style="color:#05BAC3"> Part 1  Safety Level Rating System</strong>
 +
  <br><br>
 +
</div>
 +
<div class="p">
 +
  <p>In China, the Biosafety laboratory means that laboratory structures and facilities, safety practices and safety equipment can ensure that workers are not contaminated by subjects infected with pathogenic microorganisms and toxins, and that the surrounding environment is unaffected. According to the harm degree of microorganism and toxin, it is divided into four levels with the lowest level of one and the highest level of four. And they are named after the safety standards that are required to equip and experiment: BSL-1 to BSL-4.</p>
 +
  <p> Biosafety-level 1 laboratories are generally applicable to microorganisms that have no pathogenic effect on healthy adults; the second level is applicable to microorganisms with moderate potential hazards to humans and the environment; the third level is applicable to pathogenic microorganisms or toxins that cause severe or even fatal diseases, mainly through respiratory pathways; level four is applicable to the human body with a high degree of risk, and the pathogenic microorganism or its toxin, which is unknown and has no effective vaccine or treatment method, is transmitted or transmitted by means of a vapor-sol pathway.    </p>
 +
  <p>With the friendly allowance of the Institution of Molecular Biology and Biological Chemistry (Prof. Yuan and Prof. Ren) and the Open and Creativity Laboratory (Prof. Cao), we are able to conduct our research at Shandong University. During our experiment, we received friendly help and instruction from professors and graduate students there so that we can finish our plan as scheduled successfully. They gave us some guidance and helped us analyze how we did, which made our work more efficient and fruitful. </p>
 +
</div>
 +
<div class="img"><img src="https://static.igem.org/mediawiki/2017/d/d0/Part01.png" style="width:30%;height:auto;margin-left:2em;">
 +
<img src="https://static.igem.org/mediawiki/2017/3/30/Part02.png" style="width:30%;height:auto;margin-left:4em;"></div>
 +
</div>
 +
<br><br>
  
 +
<div>
 +
<div class="title" >
 +
<strong style="color:#D5292C"> Part 2 General laboratory safety</strong>
 +
  </div>
 +
<div class="p">
 +
  <p>All work were conducted in the BSL-2 laboratory, which required additional training and safety measures. Before entering the laboratory, we have been trained for several days. We received training from professional laboratory manager Prof. Cao and Prof. Ren. And we learned the standards of laboratory safety (See more in the attachment and Part 3).</p>
 +
  <p>&nbsp;</p>
 +
</div>
 +
<div class="img"><img src="https://static.igem.org/mediawiki/2017/a/ab/Part03.png" style="width:30%;height:auto;margin-left:2em;">
 +
<img src="https://static.igem.org/mediawiki/2017/b/b1/Part04.png" style="width:30%;height:auto;margin-left:4em;"></div>
 
</div>
 
</div>
 +
<br><br><br><br><br><br><br><br>
 +
<br><br><br><br>
  
 +
  
<div class="column full_size">
 
<h5>Safe Project Design</h5>
 
  
<p>Does your project include any safety features? Have you made certain decisions about the design to reduce risks? Write about them here! For example:</p>
+
<div>
 
+
<div class="title" >
<ul>
+
<strong style="color:#D5292C">Part 3 Work condition and environmentt</strong>
<li>Choosing a non-pathogenic chassis</li>
+
  </div>
<li>Choosing parts that will not harm humans / animals / plants</li>
+
<div class="p">
<li>Substituting safer materials for dangerous materials in a proof-of-concept experiment</li>
+
  <p>&nbsp;</p>
<li>Including an "induced lethality" or "kill-switch" device</li>
+
  <p>We obey the following rules of laboratory strictly: Do not eat, or smoke, clean contact lenses and make-up in the work area. Food should be stored in a special cabinet or refrigerator outside the work area. Do not use the mouth pipetting, and one can only use mechanical device pipetting. Obey standard use of sharp norms. All the operation should be as carefully as possible to avoid spill and aerosol. Use effective disinfector that is effective to pathogens when the experiment is completed and before getting off work. All cultures, reservoirs and other specified wastes should be disinfected using a viable disinfection method prior to release, such as autoclaving. Transfer to the nearest laboratory disinfection of the material should be placed in durable, leak-proof containers, sealed out of the laboratory. Dispose of the material from the system for disinfection prior to transfer, and its packaging should comply with the relevant local, state, federal regulations.</p>
</ul>
+
    <p> We have separated places for experiment and protocol. Before and after our work for a day, we cleaned the laboratory with proper methods, from desk, stools, floor to equipment such as centrifuge, clean bench, tubes and so on. We were careful when using the centrifuge to avoid production of aerosol and imbalance. We put reagent and equipment into certain places according to their characteristics. We prepared things we need for the next day. All the tubes, pipette tips, EP tubes, PCR small tubes and culture medium we used were disinfected with autoclaves. As for wastes, we divided them into regulate ones that were not harm to environment and can be disposed as normal rubbish, and laboratory ones that needed to be sterilized with an autoclave to make them harmless or special management, and the later were put into a specialized bag for laboratory waste and sent to certain institution for disposal. <p>
 +
</div>
 +
<div class="img"><img src="https://static.igem.org/mediawiki/2017/3/32/Part05.png" style="width:30%;height:auto;margin-left:2em;">
 +
<img src="https://static.igem.org/mediawiki/2017/1/11/Part06.png" style="width:30%;height:auto;margin-left:4em;"></div>
 +
</div>
 +
<br><br><br><br><br><br><br><br>
 +
<br><br><br><br>
 +
 
 +
<div>
 +
<div class="title" >
 +
  <p>&nbsp;</p>
 +
  <strong style="color:#D5292C">Part 4 Personal Protective Equipment</strong>
 +
  <p>&nbsp;</p>
 +
</div>
 +
<div class="p">
 +
  <p>We wore lab coats that are made of cotton, enclosed leather shoes and rubber inspection gloves all the time. When working with corrosives, acids, bases, and alcohols, safety glasses were also required. Upon leaving the lab, we used a disinfectant hand-wash.    </p>
 +
  <p>And there are some safety equipment, including the operation of emergency showers and eyewash, as well as other emergency equipment including a fire blanket, extinguisher, and the panic button.
 +
    While working with EtBr, we used thicker nitrile gloves and layers of plastic gloves to protect ourselves from this dangerous chemical. And any operation with EtBr were under control to ensure safety.</p>
 +
</div>
 +
</div>
 +
<br><br><br>
  
 +
<div>
 +
<div class="title" >
 +
<strong style="color:#D5292C">Part 5 Organisms and parts that were used</strong>
 +
  <p>&nbsp;</p>
 
</div>
 
</div>
 +
<div class="STYLE1">
 +
  <p>Organisms</p>
 +
  </div>
 +
  <table width="80%" border="1">
 +
  <tr>
 +
    <th scope="col"><p >Species name (including strain)</p></th>
 +
    <th scope="col"><p >Risk Group</p></th>
 +
    <th scope="col"><p >Risk Group Source</p></th>
 +
    <th scope="col"><p >Disease risk to humans?</p></th>
 +
  </tr>
 +
  <tr>
 +
    <td width="202" valign="top" ><p align="center" >E.coli DH-5&#945; </p></td>
 +
    <td width="88" valign="top" ><p align="center" >1 </p></td>
 +
    <td width="141" valign="top" ><p align="center" >NIH </p></td>
 +
    <td width="198" valign="top" ><p align="center" >No </p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="202" valign="top" ><p align="center" >H460 </p></td>
 +
    <td width="88" valign="top" ><p align="center" >2 </p></td>
 +
    <td width="141" valign="top" ><p align="center" >NIH </p></td>
 +
    <td width="198" valign="top" ><p align="center" >No </p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="202" valign="top" ><p align="center" >A549 </p></td>
 +
    <td width="88" valign="top" ><p align="center" >2 </p></td>
 +
    <td width="141" valign="top" ><p align="center" >NIH </p></td>
 +
    <td width="198" valign="top" ><p align="center" >No </p></td>
 +
  </tr>
 +
  <tr>
 +
    <td width="202" valign="top" ><p align="center" >293T </p></td>
 +
    <td width="88" valign="top" ><p align="center" >1 </p></td>
 +
    <td width="141" valign="top" ><p align="center" >NIH </p></td>
 +
    <td width="198" valign="top" ><p align="center" >No </p></td>
 +
  </tr>
 +
</table>
  
<div class="column half_size">
+
<div class="subtitle">
<h5>Safe Lab Work</h5>
+
  <p>&nbsp;</p>
 
+
  <p><strong>Parts</strong></p>
<p>What safety procedures do you use every day in the lab? Did you perform any unusual experiments, or face any unusual safety issues? Write about them here!</p>
+
 
+
 
</div>
 
</div>
 +
<table width="80%" border="1">
 +
  <tr >
 +
    <td valign="center" ><p align="center" >Part number/name </p></td>
 +
    <td valign="center" ><p align="center" >Natural function of part </p></td>
 +
    <td valign="center" ><p align="center" >How did you acquire it? </p></td>
 +
    <td valign="center" ><p align="center" >How will you use it? </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >pSB1C3 - BBa_J04500 </p></td>
 +
    <td valign="center" ><p align="center" >IPTG inducible promoter with RBS </p></td>
 +
    <td valign="center" ><p align="center" >We acquired it from iGEM DNA Distribution Kits. </p></td>
 +
    <td valign="center" ><p align="center" >Transcription in prokaryotic bacteria </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >pSB1C3 - BBa_B0015 </p></td>
 +
    <td valign="center" ><p align="center" >Cause transcription termination </p></td>
 +
    <td valign="center" ><p align="center" >We acquired it from iGEM DNA Distribution Kits. </p></td>
 +
    <td valign="center" ><p align="center" > Transcription termination </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >pSB1C3 - BBa_K592012 </p></td>
 +
    <td valign="center" ><p align="center" >Coding red chromoprotein </p></td>
 +
    <td valign="center" ><p align="center" >We acquired it from iGEM DNA Distribution Kits. </p></td>
 +
    <td valign="center" ><p align="center" >As a reporter gene </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >pSB1C3 - BBa_E0840 </p></td>
 +
    <td valign="center" ><p align="center" >GFP generator </p></td>
 +
    <td valign="center" ><p align="center" >We acquired it from iGEM DNA Distribution Kits. </p></td>
 +
    <td valign="center" ><p align="center" >As a reporter gene </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >Promoter 1(hTERT) </p></td>
 +
    <td valign="center" ><p align="center" >Start up plasmid expression in toumor cell </p></td>
 +
    <td valign="center" ><p align="center" > Order the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >Start up plasmid expression in toumor cell </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >Promoter 2(Survivin) </p></td>
 +
    <td valign="center" ><p align="center" >Start up plasmid expression in toumor cell </p></td>
 +
    <td valign="center" ><p align="center" > Ordered the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >Start up plasmid expression in toumor cell </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >tetR </p></td>
 +
    <td valign="center" ><p align="center" >Express tetR protein </p></td>
 +
    <td valign="center" ><p align="center" > Order the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >Used it to inhibited the function of tetR repressible promoter </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >tetR represible promoter </p></td>
 +
    <td valign="center" ><p align="center" >It can't start up the expression of plasmid when it combine with tetR protein. </p></td>
 +
    <td valign="center" ><p align="center" > Order the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >Play a role of regulation </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >amilCP </p></td>
 +
    <td valign="center" ><p align="center" >Coding purple protein </p></td>
 +
    <td valign="center" ><p align="center" > Order the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >AS a reporter gene </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >eforRed </p></td>
 +
    <td valign="center" ><p align="center" >Coding red protein </p></td>
 +
    <td valign="center" ><p align="center" > Order the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >AS a reporter gene </p></td>
 +
  </tr>
 +
  <tr >
 +
    <td valign="center" ><p align="center" >neomycin </p></td>
 +
    <td valign="center" ><p align="center" >Express protein to resistant neomycin </p></td>
 +
    <td valign="center" ><p align="center" > Order the part DNA from a synthesis company </p></td>
 +
    <td valign="center" ><p align="center" >For screening </p></td>
 +
  </tr>
 +
</table>
 +
<br><br><br>
  
<div class="column half_size">
+
<div>
<h5>Safe Shipment</h5>
+
<div class="title" >
 
+
  <p><strong style="color:#D5292C">Part 6 Risk assessment</strong></p>
<p>Did you face any safety problems in sending your DNA parts to the Registry? How did you solve those problems?</p>
+
  </div>
 +
<div class="p">
 +
  <p>As our project is to use gene therapy as the heart to conquer non-small cells lung cancer (NSCLC), and we also hope this kind of treatments can serve as a weapon used in real clinical treatment someday, so we are very concerned about the safety of plasmid vectors we constructed when they are delivered into cells.    </p>
 +
  <p>We believe that risks are mainly embodied in two aspects. The first one is the efficiency of our system based on the major premise plasmids triger gene expression in cells. So the sensitivity of the tumor specific promoter and the specificity of the sgRNA is fundamental. The other one is that plamids could spread through not only the tumor cells but also the normal cells, which contains potential risks.</p>
 +
  <p> In order to test the working condition of the whole system, validation was performed for every part. In the experiment, we have inserted the hTERT and Survivin promoters respectively to pgl-3 vectors containing double luciferases reporter genes and then transferred the constructed plasmids into cancer cells as well as normal cells respectively, and finally measured the luciferase activity of each group, which could indicated the sensitivity of the tumor specific promoters. Meanwhile, various sgRNA was connected to LentiCRISPR v2 which could be transferred into NSCLC cells and normal cells, and we could detect the protein of PD-L1 through PCR and endonuclease BsmBI. In this way, best promoter and sgRNA were chosen already.
 +
    To further reduce the unpredictable of the plasmids flow, we design another knockout system based on the technique of CRISPR/Cas9, minimizing the effect of the plasmids. The concrete design is that when the plasmids are transferred into normal cells that hTERT/Survivin promoter can not start gene expression, two plasmids would achieve suicide to ensure biosafety via the sgRNA targeting the gene of Cas9. </p>
 
</div>
 
</div>
 +
<div class="img"><img src="https://static.igem.org/mediawiki/2017/b/b4/Part07.png" style="width:60%;height:auto;margin-left:2em;"></div>
 +
</div>
 +
<br><br><br><br><br><br>
  
 
+
<a href="https://static.igem.org/mediawiki/2017/2/21/Rules.pdf"/><p>Click on and download the safety rules</p><a>
 +
</div>
 +
</body>
 
</html>
 
</html>

Revision as of 05:28, 22 October 2017






Part 1 Safety Level Rating System

In China, the Biosafety laboratory means that laboratory structures and facilities, safety practices and safety equipment can ensure that workers are not contaminated by subjects infected with pathogenic microorganisms and toxins, and that the surrounding environment is unaffected. According to the harm degree of microorganism and toxin, it is divided into four levels with the lowest level of one and the highest level of four. And they are named after the safety standards that are required to equip and experiment: BSL-1 to BSL-4.

Biosafety-level 1 laboratories are generally applicable to microorganisms that have no pathogenic effect on healthy adults; the second level is applicable to microorganisms with moderate potential hazards to humans and the environment; the third level is applicable to pathogenic microorganisms or toxins that cause severe or even fatal diseases, mainly through respiratory pathways; level four is applicable to the human body with a high degree of risk, and the pathogenic microorganism or its toxin, which is unknown and has no effective vaccine or treatment method, is transmitted or transmitted by means of a vapor-sol pathway.

With the friendly allowance of the Institution of Molecular Biology and Biological Chemistry (Prof. Yuan and Prof. Ren) and the Open and Creativity Laboratory (Prof. Cao), we are able to conduct our research at Shandong University. During our experiment, we received friendly help and instruction from professors and graduate students there so that we can finish our plan as scheduled successfully. They gave us some guidance and helped us analyze how we did, which made our work more efficient and fruitful.



Part 2 General laboratory safety

All work were conducted in the BSL-2 laboratory, which required additional training and safety measures. Before entering the laboratory, we have been trained for several days. We received training from professional laboratory manager Prof. Cao and Prof. Ren. And we learned the standards of laboratory safety (See more in the attachment and Part 3).

 













Part 3 Work condition and environmentt

 

We obey the following rules of laboratory strictly: Do not eat, or smoke, clean contact lenses and make-up in the work area. Food should be stored in a special cabinet or refrigerator outside the work area. Do not use the mouth pipetting, and one can only use mechanical device pipetting. Obey standard use of sharp norms. All the operation should be as carefully as possible to avoid spill and aerosol. Use effective disinfector that is effective to pathogens when the experiment is completed and before getting off work. All cultures, reservoirs and other specified wastes should be disinfected using a viable disinfection method prior to release, such as autoclaving. Transfer to the nearest laboratory disinfection of the material should be placed in durable, leak-proof containers, sealed out of the laboratory. Dispose of the material from the system for disinfection prior to transfer, and its packaging should comply with the relevant local, state, federal regulations.

We have separated places for experiment and protocol. Before and after our work for a day, we cleaned the laboratory with proper methods, from desk, stools, floor to equipment such as centrifuge, clean bench, tubes and so on. We were careful when using the centrifuge to avoid production of aerosol and imbalance. We put reagent and equipment into certain places according to their characteristics. We prepared things we need for the next day. All the tubes, pipette tips, EP tubes, PCR small tubes and culture medium we used were disinfected with autoclaves. As for wastes, we divided them into regulate ones that were not harm to environment and can be disposed as normal rubbish, and laboratory ones that needed to be sterilized with an autoclave to make them harmless or special management, and the later were put into a specialized bag for laboratory waste and sent to certain institution for disposal.













 

Part 4 Personal Protective Equipment

 

We wore lab coats that are made of cotton, enclosed leather shoes and rubber inspection gloves all the time. When working with corrosives, acids, bases, and alcohols, safety glasses were also required. Upon leaving the lab, we used a disinfectant hand-wash.

And there are some safety equipment, including the operation of emergency showers and eyewash, as well as other emergency equipment including a fire blanket, extinguisher, and the panic button. While working with EtBr, we used thicker nitrile gloves and layers of plastic gloves to protect ourselves from this dangerous chemical. And any operation with EtBr were under control to ensure safety.




Part 5 Organisms and parts that were used

 

Organisms

Species name (including strain)

Risk Group

Risk Group Source

Disease risk to humans?

E.coli DH-5α

1

NIH

No

H460

2

NIH

No

A549

2

NIH

No

293T

1

NIH

No

 

Parts

Part number/name

Natural function of part

How did you acquire it?

How will you use it?

pSB1C3 - BBa_J04500

IPTG inducible promoter with RBS

We acquired it from iGEM DNA Distribution Kits.

Transcription in prokaryotic bacteria

pSB1C3 - BBa_B0015

Cause transcription termination

We acquired it from iGEM DNA Distribution Kits.

 Transcription termination

pSB1C3 - BBa_K592012

Coding red chromoprotein

We acquired it from iGEM DNA Distribution Kits.

As a reporter gene

pSB1C3 - BBa_E0840

GFP generator

We acquired it from iGEM DNA Distribution Kits.

As a reporter gene

Promoter 1(hTERT)

Start up plasmid expression in toumor cell

 Order the part DNA from a synthesis company

Start up plasmid expression in toumor cell

Promoter 2(Survivin)

Start up plasmid expression in toumor cell

 Ordered the part DNA from a synthesis company

Start up plasmid expression in toumor cell

tetR

Express tetR protein

 Order the part DNA from a synthesis company

Used it to inhibited the function of tetR repressible promoter

tetR represible promoter

It can't start up the expression of plasmid when it combine with tetR protein.

 Order the part DNA from a synthesis company

Play a role of regulation

amilCP

Coding purple protein

 Order the part DNA from a synthesis company

AS a reporter gene

eforRed

Coding red protein

 Order the part DNA from a synthesis company

AS a reporter gene

neomycin

Express protein to resistant neomycin

 Order the part DNA from a synthesis company

For screening




Part 6 Risk assessment

As our project is to use gene therapy as the heart to conquer non-small cells lung cancer (NSCLC), and we also hope this kind of treatments can serve as a weapon used in real clinical treatment someday, so we are very concerned about the safety of plasmid vectors we constructed when they are delivered into cells.

We believe that risks are mainly embodied in two aspects. The first one is the efficiency of our system based on the major premise plasmids triger gene expression in cells. So the sensitivity of the tumor specific promoter and the specificity of the sgRNA is fundamental. The other one is that plamids could spread through not only the tumor cells but also the normal cells, which contains potential risks.

In order to test the working condition of the whole system, validation was performed for every part. In the experiment, we have inserted the hTERT and Survivin promoters respectively to pgl-3 vectors containing double luciferases reporter genes and then transferred the constructed plasmids into cancer cells as well as normal cells respectively, and finally measured the luciferase activity of each group, which could indicated the sensitivity of the tumor specific promoters. Meanwhile, various sgRNA was connected to LentiCRISPR v2 which could be transferred into NSCLC cells and normal cells, and we could detect the protein of PD-L1 through PCR and endonuclease BsmBI. In this way, best promoter and sgRNA were chosen already. To further reduce the unpredictable of the plasmids flow, we design another knockout system based on the technique of CRISPR/Cas9, minimizing the effect of the plasmids. The concrete design is that when the plasmids are transferred into normal cells that hTERT/Survivin promoter can not start gene expression, two plasmids would achieve suicide to ensure biosafety via the sgRNA targeting the gene of Cas9.







Click on and download the safety rules