Difference between revisions of "Part:BBa K3738020"

 
 
(One intermediate revision by the same user not shown)
Line 1: Line 1:
  
 +
__NOTOC__
 +
<partinfo>BBa_K3738020 short</partinfo>
 +
 +
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.
 +
 +
 +
<!-- Add more about the biology of this part here
 +
===Usage and Biology===
 +
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K3738020 SequenceAndFeatures</partinfo>
 +
 +
 +
<!-- Uncomment this to enable Functional Parameter display
 +
===Functional Parameters===
 +
<partinfo>BBa_K3738020 parameters</partinfo>
 +
<!-- -->

Latest revision as of 17:04, 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 PstI site found at 1089
    Illegal PstI site found at 1944
    Illegal PstI site found at 2883
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1089
    Illegal PstI site found at 1944
    Illegal PstI site found at 2883
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 370
    Illegal BglII site found at 1306
    Illegal BglII site found at 1516
    Illegal BglII site found at 1570
    Illegal BglII site found at 1801
    Illegal BglII site found at 2074
    Illegal BglII site found at 2560
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1089
    Illegal PstI site found at 1944
    Illegal PstI site found at 2883
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1089
    Illegal PstI site found at 1944
    Illegal PstI site found at 2883
    Illegal NgoMIV site found at 2022
    Illegal NgoMIV site found at 2658
  • 1000
    COMPATIBLE WITH RFC[1000]