Difference between revisions of "Part:BBa K1323002"
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<partinfo>BBa_K1323002 short</partinfo> | <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 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 | + | 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. |
+ | This part was DNA-Synthesized. | ||
+ | |||
+ | References | ||
+ | Lei S. Qi, M. H. (2013). Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 1173-1183. | ||
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===Usage and Biology=== | ===Usage and Biology=== |
Revision as of 01:31, 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.
This part was DNA-Synthesized.
References Lei S. Qi, M. H. (2013). Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell, 1173-1183. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1476
Illegal BamHI site found at 4894 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]