Difference between revisions of "Part:BBa K1797016"
Morning ZHOU (Talk | contribs) |
|||
Line 1: | Line 1: | ||
− | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1797016 short</partinfo> | <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. | 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 | <!-- Add more about the biology of this part here |
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.
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:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1099
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3378
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]