Difference between revisions of "Part:BBa K2130000"
Line 7: Line 7:
  
  
−
<html><center><img src="https://static.igem.org/mediawiki/2016/0/0b/T--NTU-Singapore--ruvc3.jpg" height="60%"width="75%">
+
<html><center><img src="https://static.igem.org/mediawiki/2016/0/0b/T--NTU-Singapore--ruvc3.jpg" height="60%"width="75%"><p>Conformation of dCas9 before and after non-targeted strand cleavage.</p>
  
 
<img src="https://static.igem.org/mediawiki/2016/a/a2/T--NTU-Singapore--ruvc3delresult.png" height="60%"width="75%">
 
<img src="https://static.igem.org/mediawiki/2016/a/a2/T--NTU-Singapore--ruvc3delresult.png" height="60%"width="75%">
  
−
<p>Binding Affinity relative to other truncated dCas9s.</p>
+
 
  
 
<img src="https://static.igem.org/mediawiki/2016/e/e2/T--NTU-Singapore--conclusion.png" height="60%"width="75%">
 
<img src="https://static.igem.org/mediawiki/2016/e/e2/T--NTU-Singapore--conclusion.png" height="60%"width="75%">

Revision as of 14:48, 22 October 2016


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


Conformation of dCas9 before and after non-targeted strand cleavage.


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 1188
    Illegal BamHI site found at 2006
    Illegal XhoI site found at 2962
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2350
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2496
    Illegal BsaI site found at 3158
    Illegal BsaI.rc site found at 1411