Part:BBa_J428061
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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2830
Illegal PstI site found at 2851 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2830
Illegal PstI site found at 2851 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2830
Illegal PstI site found at 2851 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2830
Illegal PstI site found at 2851 - 1000COMPATIBLE WITH RFC[1000]
- ↑ 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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