Difference between revisions of "Part:BBa K786031:Design"

 
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===Design Notes===
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<strong>I. Mechanism: CRISPR/Cas systems</strong>
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The repeat-spacer-repeat region design should be consistent with the operating principles of E.coli's CRISPR/cas system.
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  <p>CRISPR/Cas systems are the adaptive immunity systems that are present in many archaea and bacteria to protect themselves from invading genetic materials such as viral DNA. There are three types of CRISPR/Cas system. Here we only exploit type I-E from <em>Escherichia coli</em> (<em>E. coli</em>) to focus on expression and interference stage. This system presents potential in developing into a novel tool in order to reach higher safety standard of using engineered microorganism machine.<br />
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        Foreign double strand DNA (dsDNA) will be targeted by the spacers from repeat-spacer region (RSR) and destroyed by one of the CRISPR/Cas components, Cas3. Through engineering the spacer sequence, theoretically we can target any dsDNA of sequence complementary to the spacer(s) [1].<br /><br />
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        By re-designing the targeting sequence, we can cleave the transformed plasmid in certain bacteria to reduce environmental hazard, or even manage to create a controlled suicide system to reduce leakage of genetic information from the bacteria. Thus our system provides an alternative to control safety level of engineered microorganism machine.  
  
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II. <strong>Biobrick design</strong>
  
 
===Source===
 
===Source===

Revision as of 17:13, 27 September 2012

T7promoter-RSR-T7teminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 816
  • 1000
    COMPATIBLE WITH RFC[1000]


I. Mechanism: CRISPR/Cas systems

CRISPR/Cas systems are the adaptive immunity systems that are present in many archaea and bacteria to protect themselves from invading genetic materials such as viral DNA. There are three types of CRISPR/Cas system. Here we only exploit type I-E from Escherichia coli (E. coli) to focus on expression and interference stage. This system presents potential in developing into a novel tool in order to reach higher safety standard of using engineered microorganism machine.

Foreign double strand DNA (dsDNA) will be targeted by the spacers from repeat-spacer region (RSR) and destroyed by one of the CRISPR/Cas components, Cas3. Through engineering the spacer sequence, theoretically we can target any dsDNA of sequence complementary to the spacer(s) [1].

By re-designing the targeting sequence, we can cleave the transformed plasmid in certain bacteria to reduce environmental hazard, or even manage to create a controlled suicide system to reduce leakage of genetic information from the bacteria. Thus our system provides an alternative to control safety level of engineered microorganism machine.
II. Biobrick design

Source

The 'repeat' sequence in CRISPR/cas system of E.coli strain K12.

===References===