Difference between revisions of "Part:BBa K3982008:Design"

 
 
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===Design Notes===
 
===Design Notes===
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None
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The scaffold region has been designed as referred from Ref. 1. It consists of the tracrRNA and some regions of crRNA. The guiding region is the 20 bp spacer sequence which is complementary to the mutant rpoB gene in Mtb and referred from Ref 2. The mutation appears at base number 11 (tcg → ttg)  of the spacer (guiding) sequence. This is the S450L mutation in Mtb genome (refer Ref 3).
  
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Refer to the wildtype katG targeting sgRNA here  -  [https://parts.igem.org/Part:BBa_K3982007 BBa_K3982007]
  
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The 20bp sequence is as follows: cgccgactgttggcgctggg
  
 
===Source===
 
===Source===
  
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NCBI Reference Sequence: NC_000962.3
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===References===
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Reference: L. B. Harrington et al. Science 10.1126/science.aav4294 (2018)
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1) Kim, D.Y., Lee, J.M., Moon, S.B. et al. Efficient CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs delivered by adeno-associated virus. Nat Biotechnol (2021). https://doi.org/10.1038/s41587-021-01009-z
  
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2) NCBI Reference Sequence for Mycobacterium tuberculosis H37Rv: NC_000962.3
  
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===References===
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3) Kumar, S., & Jena, L. (2014). Understanding Rifampicin Resistance in Tuberculosis through a Computational Approach. Genomics & informatics, 12(4), 276–282. https://doi.org/10.5808/GI.2014.12.4.276

Latest revision as of 13:04, 11 October 2021


CODE M sgRNA for targeting mutated rpoB gene in Mycobacterium tuberculosis


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

The scaffold region has been designed as referred from Ref. 1. It consists of the tracrRNA and some regions of crRNA. The guiding region is the 20 bp spacer sequence which is complementary to the mutant rpoB gene in Mtb and referred from Ref 2. The mutation appears at base number 11 (tcg → ttg) of the spacer (guiding) sequence. This is the S450L mutation in Mtb genome (refer Ref 3).

Refer to the wildtype katG targeting sgRNA here - BBa_K3982007

The 20bp sequence is as follows: cgccgactgttggcgctggg

Source

References

1) Kim, D.Y., Lee, J.M., Moon, S.B. et al. Efficient CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs delivered by adeno-associated virus. Nat Biotechnol (2021). https://doi.org/10.1038/s41587-021-01009-z

2) NCBI Reference Sequence for Mycobacterium tuberculosis H37Rv: NC_000962.3

3) Kumar, S., & Jena, L. (2014). Understanding Rifampicin Resistance in Tuberculosis through a Computational Approach. Genomics & informatics, 12(4), 276–282. https://doi.org/10.5808/GI.2014.12.4.276