Coding

Part:BBa_K1699003:Design

Designed by: Emil Ruvinov   Group: iGEM15_BGU_Israel   (2015-08-09)
Revision as of 11:13, 13 September 2015 by Emilr78 (Talk | contribs) (→‎Design Notes)

gRNA for dCas9-VP64 targeting synthetic activation promoter pMLPm


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 144
    Illegal NgoMIV site found at 173
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Designed using Benchling to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restrictoion sites.

Source

Sequence: Tunable and multifunctional eukaryotic transcription factors based on CRISPR/Cas. Farzadfard F, Perli SD, Lu TK. ACS Synth Biol. 2013 Oct 18;2(10):604-13. doi: 10.1021/sb400081r. Epub 2013 Sep 11. http://www.ncbi.nlm.nih.gov/pubmed/23977949

Ribozyme-flanked (RGR) design for the gRNA synthesis under RNA polymerase II promoters: Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing. Gao Y, Zhao Y. J Integr Plant Biol. 2014 Apr;56(4):343-9. doi: 10.1111/jipb.12152. Epub 2014 Mar 6. http://www.ncbi.nlm.nih.gov/pubmed/24373158 Multiplexed and programmable regulation of gene networks with an integrated RNA and CRISPR/Cas toolkit in human cells. Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK. Mol Cell. 2014 May 22;54(4):698-710. doi: 10.1016/j.molcel.2014.04.022. Epub 2014 May 15. http://www.ncbi.nlm.nih.gov/pubmed/24837679

References

1. Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing
http://onlinelibrary.wiley.com/doi/10.1111/jipb.12152/full

2. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
http://pubs.acs.org/doi/full/10.1021/sb400081r

3. In vivo genome editing using Staphylococcus aureus Cas9.
http://10.1038/nature14299