Difference between revisions of "Part:BBa K512000"

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===Usage and Biology===
 
===Usage and Biology===
 
cas3 belongs to a series of genes responsible for CRISPR interference to work. cas3 has been described as a protein essential to the functionality of the Cascade (CRISPR-associated complex for antiviral defence) complex. The cas3 protein is believed to be a helicase that assists mature CRISPR transcripts (RNA) to enable interference with virus proliferation in vivo.
 
cas3 belongs to a series of genes responsible for CRISPR interference to work. cas3 has been described as a protein essential to the functionality of the Cascade (CRISPR-associated complex for antiviral defence) complex. The cas3 protein is believed to be a helicase that assists mature CRISPR transcripts (RNA) to enable interference with virus proliferation in vivo.
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Proof of operation:
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[[Image:Cas3 function.jpg]]
 
[[Image:Cas3 function.jpg]]
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[http://2011.igem.org/Team:USC/Project]
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In the chart above, BL-D1 are the cells without IPTG, BL-D2 are with IPTG; and for BL-D3, we add IPTG 2 hours later. IPTG can induce CRISPR to target the plasmid. As we can see from the chart, with cas3 and CRISPR, the growth rate and optical density are decreased significantly  compare to the cells without IPTG.
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For complete and other results, please check the link [http://2011.igem.org/Team:USC/Project]
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 02:49, 29 September 2011

cas3

Usage and Biology

cas3 belongs to a series of genes responsible for CRISPR interference to work. cas3 has been described as a protein essential to the functionality of the Cascade (CRISPR-associated complex for antiviral defence) complex. The cas3 protein is believed to be a helicase that assists mature CRISPR transcripts (RNA) to enable interference with virus proliferation in vivo.

Proof of operation:


Cas3 function.jpg


In the chart above, BL-D1 are the cells without IPTG, BL-D2 are with IPTG; and for BL-D3, we add IPTG 2 hours later. IPTG can induce CRISPR to target the plasmid. As we can see from the chart, with cas3 and CRISPR, the growth rate and optical density are decreased significantly compare to the cells without IPTG.

For complete and other results, please check the link [http://2011.igem.org/Team:USC/Project]

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1964
    Illegal PstI site found at 2302
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1581
    Illegal PstI site found at 1964
    Illegal PstI site found at 2302
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1964
    Illegal PstI site found at 2302
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1964
    Illegal PstI site found at 2302
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 214
    Illegal SapI site found at 904


Source

cas3 has been cloned from genomic DNA of DH5a.

References

  1. Brouns, S.J.J., et. al. (2008) Small CRISPR RNAs guide antiviral defense in prokaryotes. Science 321: 960-964.