Difference between revisions of "Part:BBa K1323002"

Revision as of 17:47, 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 dCas9 simply binds to it. This means dCas9 can function as a repressor after complexing with sgRNA (BBa_K1323000, BBa_K1323001) and the sgRNA-target site. dCas9 contains mutations in the RuvC1 and HNH nuclease domains; these changes are what bestow this modified Cas9 with its ability to bind DNA instead of cleaving it (Qi et al., 2013). BBa_K1323002 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.

This part was DNA-synthesized by Bio Basic.

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]

References

Qi, L.S. et al., (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell, 1173-1183.

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1323002 short</partinfo>
-dCas9 or ‘dead’ cas9 is a mutated version of Cas9 that has lost its endonuclease activity - it no longer cuts double stranded DNA, instead dCas9 simply binds to it. This means dCas9 can function as a repressor after complexing with sgRNA ([https://parts.igem.org/Part:BBa_K1323000 BBa_K1323000], [https://parts.igem.org/Part:BBa_K1323001 BBa_K1323001]) and the sgRNA-target site. dCas9 contains mutations in the RuvC1 and HNH nuclease domains; these changes are what bestow this modified Cas9 with its ability to bind DNA instead of cleaving it (Lei S. Qi, 2013). BBa_K1323002 has a Xylose Inducible Promoter [https://parts.igem.org/Part:BBa_K1323014 BBa_K1323014] and a ''sodA'' RBS [https://parts.igem.org/Part:BBa_K1323022 BBa_K1323022]. The illegal sites have been removed, and the sequence was codon optimized for expression in ''S.epidermidis'' using JCat software.
+dCas9 or ‘dead’ cas9 is a mutated version of Cas9 that has lost its endonuclease activity - it no longer cuts double stranded DNA, instead dCas9 simply binds to it. This means dCas9 can function as a repressor after complexing with sgRNA ([https://parts.igem.org/Part:BBa_K1323000 BBa_K1323000], [https://parts.igem.org/Part:BBa_K1323001 BBa_K1323001]) and the sgRNA-target site. dCas9 contains mutations in the RuvC1 and HNH nuclease domains; these changes are what bestow this modified Cas9 with its ability to bind DNA instead of cleaving it (Qi et al., 2013). BBa_K1323002 has a Xylose Inducible Promoter [https://parts.igem.org/Part:BBa_K1323014 BBa_K1323014] and a ''sodA'' RBS [https://parts.igem.org/Part:BBa_K1323022 BBa_K1323022]. The illegal sites have been removed, and the sequence was codon optimized for expression in ''S.epidermidis'' using JCat software.
 This part was DNA-synthesized by Bio Basic.
@@ Line 17: / Line 17: @@
 ==References==
-Lei S. Qi, M. H. (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. ''Cell'', 1173-1183.
+Qi, L.S. et al., (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. ''Cell'', 1173-1183.
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