Coding

Part:BBa_J428061

Designed by: Vinoo Selvarajah   Group: 2021 Engineering Committee   (2022-06-07)


Cas_variants_hLwCas13a

Cas_variants_hLwCas13a, synthesised from Leptotrichia wadeii


Information contributed by City of London UK (2022)

Part information is collated here to help future users of the BioBrick registry.

Metadata:

  • Group: City of London UK 2022
  • Author: Julian Chen
  • Summary: Added information collated from existing scientific studies

Usage and Biology

Cas13 is a programmable endonuclease that targets RNA, as opposed to DNA, used in Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems. It is activated by a ssRNA sequence complementary to its cRNA spacer, which then induces non-specific RNase activity that cleaves all nearby RNA sequences. It may be used for efficient and specific RNA knockdown and RNA sequencing in mammalian cells. [1]

Cas13a was considered for use by the City of London iGEM 2022 Team, alongside Cas12a, but was ultimately dropped due to the need to cleave amplified DNA sequences.

It utilises a PAM: 3’-H


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2830
    Illegal PstI site found at 2851
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2830
    Illegal PstI site found at 2851
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2830
    Illegal BglII site found at 1286
    Illegal BglII site found at 1496
    Illegal BglII site found at 1763
    Illegal BglII site found at 2528
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2830
    Illegal PstI site found at 2851
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2830
    Illegal PstI site found at 2851
  • 1000
    COMPATIBLE WITH RFC[1000]


  1. Pawluk, A., n.d. CRISPR Systems: What’s the Difference?. [online] Cell.com. Available at: <https://www.cell.com/pb-assets/products/research-arc/infographics/CrisprVizInfo_vol1a-1539275242767.pdf> [Accessed 9 October 2022].
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