Difference between revisions of "Part:BBa K2316000"
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dCas9 is a catalytically inactive Cas9, which still retains it's ability to bind to DNA. For epigenetic regulation, the dCas9 is highly dependent upon the function of it's fusion partner. This is an improvement from part <html><a href="https://parts.igem.org/Part:BBa_K2130000">BBa_K2130000</a></html>
 
dCas9 is a catalytically inactive Cas9, which still retains it's ability to bind to DNA. For epigenetic regulation, the dCas9 is highly dependent upon the function of it's fusion partner. This is an improvement from part <html><a href="https://parts.igem.org/Part:BBa_K2130000">BBa_K2130000</a></html>
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In our attempt to improve our truncations, we explored 2 possible avenues of improvements. First, we considered if further truncations is possible, by looking at additional combinations and novel truncations. Second, we considered alternative functionalities for our truncated dCas9.
  
  

Revision as of 12:45, 1 November 2017


∆RuvCIII-2 ∆HNH ∆REC2 Sp-dCas9

dCas9 is a catalytically inactive Cas9, which still retains it's ability to bind to DNA. For epigenetic regulation, the dCas9 is highly dependent upon the function of it's fusion partner. This is an improvement from part BBa_K2130000

In our attempt to improve our truncations, we explored 2 possible avenues of improvements. First, we considered if further truncations is possible, by looking at additional combinations and novel truncations. Second, we considered alternative functionalities for our truncated dCas9.


Usage and Biology

For this part, we have deleted the HNH domain, Rec2 domain and the RuvCIII-2 region of the SP-dCas9 and tested it's binding capability by assessing the strength in transcriptional activation via by tagging it with a VP64-p65-Rta tripartite activator.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 251
    Illegal BglII site found at 804
    Illegal BamHI site found at 1622
    Illegal XhoI site found at 2578
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1966
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2112
    Illegal BsaI site found at 2774
    Illegal BsaI.rc site found at 1027