Difference between revisions of "Part:BBa K1797015"

 
 
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<partinfo>BBa_K1797015 short</partinfo>
 
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In this part, we fuse dCas9(BBa_K1797006) and catalytic domain of Bxb1 gp35(BBa_K907000) with a long linker(BBa_K243006). Bxb1 gp35 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 Bxb1 gp35 is going to function is decided by gRNA.
 
In this part, we fuse dCas9(BBa_K1797006) and catalytic domain of Bxb1 gp35(BBa_K907000) with a long linker(BBa_K243006). Bxb1 gp35 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 Bxb1 gp35 is going to function is decided by gRNA.
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[[File:Serinerec.png|400px|thumb|center|Fig.1 Principles of serine 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)]]
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Bxb1 gp35 is a serine recombinase, which functions through a tetramer(Fig.1). That means when we use dcas9-bxb1 gp35 fusion protein, we need to design four sgRNAs each time, which significantly increases the specificity.
  
 
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Latest revision as of 16:47, 18 September 2015

dCas9-Long Linker (Gly-Gly-Ser-Gly)x3-Bxb1 gp35

In this part, we fuse dCas9(BBa_K1797006) and catalytic domain of Bxb1 gp35(BBa_K907000) with a long linker(BBa_K243006). Bxb1 gp35 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 Bxb1 gp35 is going to function is decided by gRNA.

Fig.1 Principles of serine 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)

Bxb1 gp35 is a serine recombinase, which functions through a tetramer(Fig.1). That means when we use dcas9-bxb1 gp35 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
    Illegal NheI site found at 4349
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3378
    Illegal BamHI site found at 4623
    Illegal XhoI site found at 4710
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 5262
    Illegal NgoMIV site found at 5349
    Illegal AgeI site found at 4399
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5457