Difference between revisions of "Part:BBa K1797016"

Latest revision as of 16:39, 18 September 2015

dCas9-Long Linker (Gly-Gly-Ser-Gly)x3-Flp

In this part, we fuse dCas9(BBa_K1797006) and catalytic domain of Flp(BBa_K1015001) with a long linker(BBa_K243006). Flp is a site-specific recombinase. Due to its specific recognition site, the use of recombinase is limited. To solve this problem, we fuse its catalytic domain with dCas9. Then where the Flp is going to function is decided by gRNA.

Fig.1 Principles of tyrosine recombinase involved recombination(source: Mechanisms of Site-Specific Recombination. Nigel D.F. Grindley, Katrine L. Whiteson, and Phoebe A. Rice. Annual Review of Biochemistry, Vol. 75: 567 -605)

Flp is a tyrosine recombinase, which functions through a tetramer(Fig.1). That means when we use dcas9-flp fusion protein, we need to design four sgRNAs each time, which significantly increases the specificity.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1099
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 3378
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K1797016 short</partinfo>
 In this part, we fuse dCas9(BBa_K1797006) and catalytic domain of Flp(BBa_K1015001) with a long linker(BBa_K243006). Flp is a site-specific recombinase. Due to its specific recognition site, the use of recombinase is limited. To solve this problem, we fuse its catalytic domain with dCas9. Then where the Flp is going to function is decided by gRNA.
+[[File:Tyrosinerecombinase.png|400px|thumb|center|Fig.1 Principles of tyrosine recombinase involved recombination(source: Mechanisms of Site-Specific Recombination. Nigel D.F. Grindley, Katrine L. Whiteson, and Phoebe A. Rice. Annual Review of Biochemistry, Vol. 75: 567 -605)]]
+Flp is a tyrosine recombinase, which functions through a tetramer(Fig.1). That means when we use dcas9-flp fusion protein, we need to design four sgRNAs each time, which significantly increases the specificity.
 <!-- Add more about the biology of this part here