Part:BBa_K2217018:Design

gRNA for Cas9 Knock-in

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

gRNA (BBa_K2217003)(Designed using MIT CRISPR web tool according to the addgene protocol).[1][2]

The most stable 2D structure of gRNA was generated using vfold[3]. The Rosetta package FARFAR[4,5] was used to build the 3D structure of gRNA.

More information on the design of this part can be found at [http://2017.igem.org/Team:AFCM-Egypt/Model# Egypt-AFCM Modeling]

Source

synthesised from gene fragments

References

[1] http://crispr.mit.edu

[2] https://media.addgene.org/cms/files/hCRISPR_gRNA_Synthesis.pdf

[3] Xu, XJ., Zhao, PN., Chen, SJ. (2014) Vfold: a web server for RNA structure and folding thermodynamics prediction.

[4] Das, R., Karanicolas, J., and Baker, D. (2010), "Atomic accuracy in predicting and designing noncanonical RNA structure". Nature Methods 7:291-294.

[5] Lyskov S, Chou FC, Conchúir SÓ, Der BS, Drew K, Kuroda D, et al. "Serverification of Molecular Modeling Applications: The Rosetta Online Server That Includes Everyone (ROSIE)". PLoS One. 2013 May 22;8(5):e63906