Difference between revisions of "Part:BBa K4140021"
Ahmed Mattar (Talk | contribs) (→Usage) |
Ahmed Wael (Talk | contribs) (→Literature Characterization) |
||
Line 13: | Line 13: | ||
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. | 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) | |
− | + | ||
+ | [[File:Acr.png|thumb|Right|Figure 1. (shows the mutational landscape of the Acr||A5 v2 anti-crisper protein.) ]] | ||
+ | |||
+ | <br><br><br><br><br><br><br> | ||
==References== | ==References== |
Revision as of 11:30, 3 October 2022
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)
References
Sequence and Features
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]