Difference between revisions of "Part:BBa K3853003"

Revision as of 21:30, 7 October 2021

dCas9

For dCas9, two nuclease splicing active domains of CRISPR associated protein 9 (Cas9), RuvC and HNH, are mutated simultaneously (D10A & H840A; RuvC- & HNH-) to obtain a protein that has no nuclease activity. The dCas9 protein has no ability to cut DNA, but can still bind to specific DNA sequences under the single-gudie RNA. The codons of the protein gene sequence are optimized to increase its expression in E.coli.

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 3576
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 3960
Illegal AgeI site found at 3304
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3853003 short</partinfo>
-A Cas9 protein with H840A and D10A mutations has only nucleic acid binding activity, but lacks nucleic acid cleavage activity
+For dCas9, two nuclease splicing active domains of CRISPR associated protein 9 (Cas9), RuvC and HNH, are mutated simultaneously (D10A & H840A; RuvC- & HNH-) to obtain a protein that has no nuclease activity. The dCas9 protein has no ability to cut DNA, but can still bind to specific DNA sequences under the single-gudie RNA. The codons of the protein gene sequence are optimized to increase its expression in <i>E.coli</i>.
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