Difference between revisions of "Part:BBa K2381004:Design"

(→‎Design Notes)
(→‎Design Notes)
 
(2 intermediate revisions by the same user not shown)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
In order to prevent the fusion protein has steric hindrance, We use a 12aa GS linker to link the light-induced dimerization protein with a part of dCas9, which split by a rational strategy of Cas9. According to the different function and spatial position between α-helix lobe and nuclease, we split dCas9 into two parts. Rad1 includes a-helix lobe, Rad2 includes nuclease lobe.
+
In order to prevent the fusion protein has steric hindrance, We use a 17aa GS linker to link the light-induced dimerization protein with a part of dCas9, which split by a rational strategy of Cas9. According to the different function and spatial position between α-helix lobe and nuclease, we split dCas9 into two parts. Rad1 includes a-helix lobe, Rad2 includes nuclease lobe. You can use this part with https://parts.igem.org/Part:BBa_K2381002
  
 
===Source===
 
===Source===
−
 
+
The light-induced protein nMagHigh is made from the VVD protein family derived from the fungus Neurospora crassa. The Rad2 is made from the dCas9 Protein derived from the Streptococcus pyogenes.
−
no
+
  
 
===References===
 
===References===
 
Wright A V, Sternberg S H, Taylor D W, et al. Rational design of a split-Cas9 enzyme complex.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(10):2984-2989.
 
Wright A V, Sternberg S H, Taylor D W, et al. Rational design of a split-Cas9 enzyme complex.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(10):2984-2989.

Latest revision as of 08:59, 27 October 2017


rad2 + nMagHigh


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1435
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 403


Design Notes

In order to prevent the fusion protein has steric hindrance, We use a 17aa GS linker to link the light-induced dimerization protein with a part of dCas9, which split by a rational strategy of Cas9. According to the different function and spatial position between α-helix lobe and nuclease, we split dCas9 into two parts. Rad1 includes a-helix lobe, Rad2 includes nuclease lobe. You can use this part with https://parts.igem.org/Part:BBa_K2381002

Source

The light-induced protein nMagHigh is made from the VVD protein family derived from the fungus Neurospora crassa. The Rad2 is made from the dCas9 Protein derived from the Streptococcus pyogenes.

References

Wright A V, Sternberg S H, Taylor D W, et al. Rational design of a split-Cas9 enzyme complex.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(10):2984-2989.