Difference between revisions of "Part:BBa K1062000"

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<partinfo>BBa_K1062000 short</partinfo>
 
<partinfo>BBa_K1062000 short</partinfo>
  
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This part is a guide RNA (gRNA) that targets RFP, when bound to Cas9. It also contains a cleavage site recognized by the Csy4 enzyme which allows it to be cleaved from neighboring DNA/RNA.
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This part is a guide RNA (gRNA) that targets the RFP gene, when bound to Cas9. It also contains a cleavage site recognized by the Csy4 enzyme which allows it to be cleaved from neighboring DNA/RNA.
 
   
 
   
  
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[[File:Conjugation project.jpg]]
 
[[File:Conjugation project.jpg]]
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In the Conjugation Project, our target contained a RFP cassette that is knocked into the chromosome of the strain. Due to the fact that the target strain is essential to proving the Conjugation Project, this gRNA is essential in order to prove that repression of a specific target can be possible.
  
 
=='''Synthetic Circuit'''==
 
=='''Synthetic Circuit'''==
  
 
[[File:Synthetic Circuit.jpg]]
 
[[File:Synthetic Circuit.jpg]]
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In the Synthetic Circuit, the target is the RFP gene that would be located in the '''High Concentration Pathway'''. The purpose of the gRNA is to prevent any possibility of the RFP gene expressing, due to a possible leaky promoter, when the '''Low Concentration Pathway''' is being induced.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 09:00, 24 September 2013

Guide RNA (gRNA) target for RFP

This part is a guide RNA (gRNA) that targets the RFP gene, when bound to Cas9. It also contains a cleavage site recognized by the Csy4 enzyme which allows it to be cleaved from neighboring DNA/RNA.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

In both of our projects, we used gRNAs and dCas9 as:

1) To specifically target and repress the "virulence gene" (we used flourescent proteins as a proof of concept) in our conjugation project

2) Improve past circuits with better decision making by using dCas9 instead of traditional repressors.

Conjugation Project

Conjugation project.jpg

In the Conjugation Project, our target contained a RFP cassette that is knocked into the chromosome of the strain. Due to the fact that the target strain is essential to proving the Conjugation Project, this gRNA is essential in order to prove that repression of a specific target can be possible.

Synthetic Circuit

Synthetic Circuit.jpg

In the Synthetic Circuit, the target is the RFP gene that would be located in the High Concentration Pathway. The purpose of the gRNA is to prevent any possibility of the RFP gene expressing, due to a possible leaky promoter, when the Low Concentration Pathway is being induced.