DNA

Part:BBa_K5412001

Designed by: Divakar Kalivarathan   Group: iGEM24_SVCE-CHENNAI   (2024-09-28)


CAS12a_S14P_S60P

This mutant CAS12a protein features specific mutations where serine residues at positions 14 and 60 have been replaced with proline. These modifications are designed to explore their effects on the enzyme's structure, activity, and specificity in CRISPR applications. This is done to improve the stability of the CAS12a enzyme required for CF detection.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1198
    Illegal PstI site found at 1696
    Illegal PstI site found at 3403
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1198
    Illegal PstI site found at 1696
    Illegal PstI site found at 3403
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1371
    Illegal BglII site found at 2775
    Illegal BglII site found at 3120
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1198
    Illegal PstI site found at 1696
    Illegal PstI site found at 3403
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1198
    Illegal PstI site found at 1696
    Illegal PstI site found at 3403
  • 1000
    COMPATIBLE WITH RFC[1000]


cas12a-mutant.png

plasmid.png Plasmid Map of CAS12A mutant in Pet-23a+

References
Zhang, L., et al. (2021). Engineering CRISPR-Cas12a for enhanced target recognition and cleavage at elevated temperatures. Nature Communications, 12, 3908. https://www.nature.com/articles/s41467-021-24117-z
Teng, F., et al. (2019). Crispr-Cas12a cleaves RNA in addition to DNA at elevated temperatures. Nature Communications, 10, 2162. https://www.nature.com/articles/s41467-019-10148-6
Swarts, D. C., et al. (2017). DNA-guided DNA interference by a prokaryotic Argonaute. Nature, 507(7491), 258-261. https://www.nature.com/articles/nature21049

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