Difference between revisions of "Part:BBa K3738020"

 
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<partinfo>BBa_K3738020 short</partinfo>
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Cas13a is an enzyme originating from Leptotrichia buccalis (Lbu) which functions to cleave single-stranded RNAs (ssRNAs); particularly mRNAs. This function is achieved following protein-RNA complex formation with CRISPR RNA (crRNA) via crRNA backbone contacts with residues from the Helical-2, HEPN1, and Linker domains of Cas13a. The crRNA contains a spacer region coding for a direct repeat stem loop as well as a region complementary to target ssRNAs. Once the enzyme complex interacts with a target ssRNA, a structural conformation change occurs within the domains of the protein that permits active site formation for non-discriminate ssRNA cleavage (O’Connel et al., 2019).
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O'Connell MR. Molecular Mechanisms of RNA Targeting by Cas13-containing Type VI CRISPR–Cas Systems. Journal of Molecular Biology. 2019;431(1):66-87. doi: https://doi.org/10.1016/j.jmb.2018.06.029.
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3738019 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K3738019 parameters</partinfo>
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Revision as of 02:56, 20 October 2021


Lbu-Cas13a with an N-Terminal 6xHistidine Tag and C-Terminal Anionic Tag

Cas13a is an enzyme originating from Leptotrichia buccalis (Lbu) which functions to cleave single-stranded RNAs (ssRNAs); particularly mRNAs. This function is achieved following protein-RNA complex formation with CRISPR RNA (crRNA) via crRNA backbone contacts with residues from the Helical-2, HEPN1, and Linker domains of Cas13a. The crRNA contains a spacer region coding for a direct repeat stem loop as well as a region complementary to target ssRNAs. Once the enzyme complex interacts with a target ssRNA, a structural conformation change occurs within the domains of the protein that permits active site formation for non-discriminate ssRNA cleavage (O’Connel et al., 2019).

O'Connell MR. Molecular Mechanisms of RNA Targeting by Cas13-containing Type VI CRISPR–Cas Systems. Journal of Molecular Biology. 2019;431(1):66-87. doi: https://doi.org/10.1016/j.jmb.2018.06.029.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7
    Illegal EcoRI site found at 868
  • 1000
    COMPATIBLE WITH RFC[1000]