Coding

Part:BBa_K4140021

Designed by: Ahmed Gamal Mohamed Mattar   Group: iGEM22_AFCM-Egypt   (2022-09-28)
Revision as of 11:30, 3 October 2022 by Ahmed Wael (Talk | contribs) (→‎Literature Characterization)


AcrIIA5 v2 anti-CRISPR


Part Description

Anti CRISPR for Cas12g Anti CRISPR are small proteins (approximately, 12–193 amino acids) which have become, quickly a new method to regulate CRISPR system activity by altering the nuclease activity or hindering binding to the target gene


Usage

We have thought to work on developing powerful protein designs to enable us to control the specificity and limit off-targeting of cas proteins. We have been relying on our protein evolution models that use deep learning to develop and predict new mutants of anti crispr proteins that are cas-specific and can act as an on-demand safety component for cas-based biosynthetic designs. This biosafety aspect is integrated with our therapeutic project for this year to develop our sensitive therapeutic design with natural brake elements (Acr-proteins) deployed to assure the safety of our design.

Characterization of Mutational Landscape

After creating a multiple sequence alignment of the protein sequence and predicting mutational landscapes, the effect of these mutations on the evolutionary fitness of the protein is tested. The prediction of the mutational landscape by saturation mutagenesis of the Acr||A5 v2 anti-crisper protein. The (R32E) mutation, as depicted in the chart, had the greatest score when compared to other mutations. On the other hand, it's clear that the (G42K) had the least evolutionary fitness for Acr||A5 v2 anti-crisper protein. As displayed in Figure(1)

A5 v2 anti-crisper protein.)








References

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 31
    Illegal EcoRI site found at 521
    Illegal XbaI site found at 25
    Illegal XbaI site found at 530
    Illegal SpeI site found at 19
    Illegal SpeI site found at 539
    Illegal PstI site found at 1
    Illegal PstI site found at 565
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 31
    Illegal EcoRI site found at 521
    Illegal SpeI site found at 19
    Illegal SpeI site found at 539
    Illegal PstI site found at 1
    Illegal PstI site found at 565
    Illegal NotI site found at 43
    Illegal NotI site found at 502
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 31
    Illegal EcoRI site found at 521
    Illegal BglII site found at 13
    Illegal BglII site found at 548
    Illegal BamHI site found at 37
    Illegal BamHI site found at 513
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 31
    Illegal EcoRI site found at 521
    Illegal XbaI site found at 25
    Illegal XbaI site found at 530
    Illegal SpeI site found at 19
    Illegal SpeI site found at 539
    Illegal PstI site found at 1
    Illegal PstI site found at 565
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 31
    Illegal EcoRI site found at 521
    Illegal XbaI site found at 25
    Illegal XbaI site found at 530
    Illegal SpeI site found at 19
    Illegal SpeI site found at 539
    Illegal PstI site found at 1
    Illegal PstI site found at 565
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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