Team:TCU Taiwan/CRISPR

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         <td><font size="3" face="Verdana" color="#333"><p>We chose CRISPR to knock-out antibiotic resistance. CRISPR system is  a bacteria immune system which is used as gene engineer machine now. It has  three types and we chose the type II CRISPR system. <br />
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         <td><font size="3" face="Verdana" color="#333"><p>We chose <a href="http://en.wikipedia.org/wiki/Phagemid" target="_blank">CRISPR</a> to knock-out antibiotic resistance. CRISPR system is  a bacteria immune system which is used as gene engineer machine now. It has  three types and we chose the type II CRISPR system. <br />
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           This system contains three parts. First, this system can express two  kinds of single strand RNA:  tracrRNA and  crRNA. crRNA&rsquo;s structure is like &ldquo;repeat---spacer---repeat&rdquo;, the spacer part is  complementary to target gene&rsquo;s DNA sequence, the repeat is complementary to  tracrRNA while tracrRNA can also have interation with a protein coded in this  system --- Cas9, an endonuclease.<br />
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           <p>This system contains three parts. First, this system can express two  kinds of single strand RNA:  tracrRNA and  crRNA. crRNA&rsquo;s structure is like &ldquo;repeat---spacer---repeat&rdquo;, the spacer part is  complementary to target gene&rsquo;s DNA sequence, the repeat is complementary to  tracrRNA while tracrRNA can also have interation with a protein coded in this  system --- Cas9, an endonuclease.</p><br />
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           Once we modified the spacer sequence of a CRISPR system and  transform it into bacteria, it will search for target gene (antibiotics resistance  gene in our project) in bacteria&rsquo;s genome by crRNA. Once the target is found,  Cas9 protein will bind on this gene&rsquo;s DNA sequence with the help of tarcrRNA  and crRNA. Then, Cas9 will make a small double strand break on the gene. So  this gene&rsquo;s sequence will be modified by later homozygous recombination, result  in the knock-out of this gene.</p></font></td>
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           <p>Once we modified the spacer sequence of a CRISPR system and  transform it into bacteria, it will search for target gene (antibiotics resistance  gene in our project) in bacteria&rsquo;s genome by crRNA. Once the target is found,  Cas9 protein will bind on this gene&rsquo;s DNA sequence with the help of tarcrRNA  and crRNA. Then, Cas9 will make a small double strand break on the gene. So  this gene&rsquo;s sequence will be modified by later homozygous recombination, result  in the knock-out of this gene.</p></font></td>
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Revision as of 15:30, 16 October 2014


 
CRISPR
 
 
CRISPR
 
Fig.1 Diagram of the possible mechanism for CRISPR.
 

We chose CRISPR to knock-out antibiotic resistance. CRISPR system is a bacteria immune system which is used as gene engineer machine now. It has three types and we chose the type II CRISPR system.

This system contains three parts. First, this system can express two kinds of single strand RNA:  tracrRNA and crRNA. crRNA’s structure is like “repeat---spacer---repeat”, the spacer part is complementary to target gene’s DNA sequence, the repeat is complementary to tracrRNA while tracrRNA can also have interation with a protein coded in this system --- Cas9, an endonuclease.


Once we modified the spacer sequence of a CRISPR system and transform it into bacteria, it will search for target gene (antibiotics resistance gene in our project) in bacteria’s genome by crRNA. Once the target is found, Cas9 protein will bind on this gene’s DNA sequence with the help of tarcrRNA and crRNA. Then, Cas9 will make a small double strand break on the gene. So this gene’s sequence will be modified by later homozygous recombination, result in the knock-out of this gene.

 
 
 
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