Difference between revisions of "Part:BBa K3044012:Design"

 
 
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<partinfo>BBa_K3044015 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3044012 SequenceAndFeatures</partinfo>
  
  
 
===Design Notes===
 
===Design Notes===
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The sgRNA was designed according to the design protocol "CRISPR interference (CRISPRi) for sequence-specific control of gene expression". [1] An important thing to keep ind mind when designing a sgRNA is off-targets. Therefore, the sgRNA sequence was aligned to the core genome of our chassis E coli K12 TOP10 and KG22 and no off-targets were found. Though, off-targets may appear in other bacteria strains.
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The sgRNA was designed according to the design protocol "CRISPR interference (CRISPRi) for sequence-specific control of gene expression". [https://www.ncbi.nlm.nih.gov/pubmed/24136345] An important thing to keep ind mind when designing a sgRNA is off-targets. Therefore, the sgRNA sequence was aligned to the core genome of our chassis E coli K12 TOP10 and KG22 and no off-targets were found. Though, off-targets may appear in other bacteria strains.
  
  

Latest revision as of 23:00, 7 October 2019


sgRNA targeting GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The sgRNA was designed according to the design protocol "CRISPR interference (CRISPRi) for sequence-specific control of gene expression". [1] An important thing to keep ind mind when designing a sgRNA is off-targets. Therefore, the sgRNA sequence was aligned to the core genome of our chassis E coli K12 TOP10 and KG22 and no off-targets were found. Though, off-targets may appear in other bacteria strains.


Source

The sequence of the sgRNA is designed on the basis of the GFP sequence. The target sequence is determined according to the location of PAM sequences in the target sequence, which the Cas protein regcognises.

References