Difference between revisions of "Part:BBa K2316001"

Revision as of 04:48, 25 October 2017

∆RuvCIII-2 ∆HNH ∆REC2 Sp-dCas9 Enhanced

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_K2130001.

Usage and Biology

For this part, we have deleted the HNH domain, Rec2 domain and the RuvCIII-2 region of the SP-dCas9. Quick changes are further done at Asn497Lys, Thr657Lys and Ser1109Arg. It was 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

@@ Line 5: / Line 5: @@
 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_K2130001.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+For this part, we have deleted the HNH domain, Rec2 domain and the RuvCIII-2 region of the SP-dCas9. Quick changes are further done at Asn497Lys, Thr657Lys and Ser1109Arg. It was tested it's binding capability by assessing the strength in transcriptional activation via by tagging it with a VP64-p65-Rta tripartite activator.
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