Difference between revisions of "Part:BBa K1699003:Design"

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===Design Notes===
 
===Design Notes===
  
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Cloned into pSBC13 using EcoRI and PstI restriction sites.
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Designed using Benchling to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restriction sites.
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<br /> F. tagtcatgGAATTCGCGGCCGCTTCTAG
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<br /> R. tgtatactCTGCAGCGGCCGCTAC
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===Source===
 
===Source===
  
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IDT de-novo synthesis.
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Sequence of gRNA:
  
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===References===
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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
  
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1. Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing
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Ribozyme-flanked (RGR) design for the gRNA synthesis under RNA polymerase II promoters:
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<br />http://onlinelibrary.wiley.com/doi/10.1111/jipb.12152/full
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2. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
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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
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<br />http://pubs.acs.org/doi/full/10.1021/sb400081r
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3. In vivo genome editing using Staphylococcus aureus Cas9.  
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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
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<br />http://10.1038/nature14299
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Latest revision as of 11:36, 13 September 2015

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 restriction sites.

Source

Sequence of gRNA:

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