Part:BBa_K512000

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:

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.