Difference between revisions of "Part:BBa K1323002"

Revision as of 00:33, 17 October 2014

dCas9: Expression cassette under a xylose inducible promoter

dCas9 or ‘dead’ cas9 is a mutated version of Cas9 that has lost its endonuclease activity - it no longer cuts double stranded DNA, instead simply binding to it. This means it can function as a repressor after complexing with sgRNA and the sgRNA-target site. dCas9 contains mutations in the RuvC1 and HNH nuclease domains; these changes are what bestow this modified Cas9 with it’s ability to bind DNA instead of cleaving it (Lei S. Qi, 2013). It has a Xylose Inducible Promoter (BBa_K1323014) and a sodA RBS (BBa_K1323022). The illegal sites have been removed, and the sequence was codon optimized for expression in S. epidermidis using JCat software.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1476
Illegal BamHI site found at 4894
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1323002 short</partinfo>
-TBA
+dCas9 or ‘dead’ cas9 is a mutated version of Cas9 that has lost its endonuclease activity - it no longer cuts double stranded DNA, instead simply binding to it. This means it can function as a repressor after complexing with sgRNA and the sgRNA-target site. dCas9 contains mutations in the RuvC1 and HNH nuclease domains; these changes are what bestow this modified Cas9 with it’s ability to bind DNA instead of cleaving it (Lei S. Qi, 2013). It has a Xylose Inducible Promoter (BBa_K1323014) and a sodA RBS (BBa_K1323022). The illegal sites have been removed, and the sequence was codon optimized for expression in S. epidermidis using JCat software.
 <!-- Add more about the biology of this part here