RNA

Part:BBa_K3982004

Designed by: Akankshya Ramkrishna Sahu   Group: iGEM21_IISER_Berhampur   (2021-10-01)
Revision as of 11:34, 11 October 2021 by Akankshya16 (Talk | contribs)


CODE M sgRNA for targeting wildtype katG gene in Mycobacterium tuberculosis

This sequence encodes for the customised sgRNA and is used for targeting wildtype katG gene in Mycobacterium tuberculosis.

Usage and Biology

Cas14a1 (also called as Un1Cas12f1) is a miniature cas protein used in CRISPR/Cas technology identified from archaebacteria and bacteriophages. It is a RNA guided nuclease which cleaves ssDNA without any Protospacer Adjacent Motif (PAM) specificity i.e. PAM independent ssDNA cleavage. This enable high-fidelity Single-nucleotide polymorphism (SNP) genotyping. It is guided by a single guide RNA (sgRNA) which has two components - CRISPR RNA (crRNA) and Trans-activating crispr RNA (tracrRNA).

crRNA is a 20bp nucleotide sequence complementary to the target DNA. tracrRNA is linked to the crRNA with a tetraloop that is responsible for cas14 activity. It plays a role in maturation of crRNA and has a stem-loop structure in itself that binds to the ribonuclease. Click hereto know more about how this part is used in project CODE M by Team IISER_Berhampur 2021.

This sgRNA has been designed specifically for targeting the wildtype katG gene along with Cas14a1 in Mycobacterium tuberculosis (Mtb) for detection of Multidrug-resistant Tuberculosis (MDR-TB).

Sequence and Features


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]


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Categories
Parameters
None