Difference between revisions of "Part:BBa K4151016"

 
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<partinfo>BBa_K4151016 short</partinfo>
 
<partinfo>BBa_K4151016 short</partinfo>
  
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The gene that expresses human activation-induced cytidine deaminase. We deleted 87 nucleotides encoding C-terminal region which contains the NES sequence to decrease the frequency of SHM.
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===Inhibit the function of AID to induce CSR===
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AID(wild type) plays a crucial role in both Somatic hypermutation(SHM) and Class switch reaction(CSR), which trigger point mutation in the variable region and constant region, respectively. With the DNA repair mechanism, double-strand breakage(DSB) repair, and nucleotide excision repair(NER), these mutation result from deamination caused by AID, leading to diversification of the antibody structure. However, in our project, we only want to utilize AID's function to induce SHM to increase the diversity of Ab's variable region. Research[1] shows that the catalytic domain of CSR is 517~594(172~198a.a). In order to inhibit AID from inducing CSR, we eliminate the C-terminus 16 a.a of AID.
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===Increase the effectiveness of SHM===
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According to a former study[2], eliminating the NES domain of AID could increase the mutation rate of SHM by up to 40 folds compared to AID(wild type). Since the catalytic domain of CSR and NES domain overlap at the C-terminus 16 a.a, eliminating the C-terminus 16 a.a can increase the efficiency of SHM while inhibiting the function of CSR.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 12:05, 11 October 2022


AID（dNES）

Inhibit the function of AID to induce CSR

AID(wild type) plays a crucial role in both Somatic hypermutation(SHM) and Class switch reaction(CSR), which trigger point mutation in the variable region and constant region, respectively. With the DNA repair mechanism, double-strand breakage(DSB) repair, and nucleotide excision repair(NER), these mutation result from deamination caused by AID, leading to diversification of the antibody structure. However, in our project, we only want to utilize AID's function to induce SHM to increase the diversity of Ab's variable region. Research[1] shows that the catalytic domain of CSR is 517~594(172~198a.a). In order to inhibit AID from inducing CSR, we eliminate the C-terminus 16 a.a of AID.

Increase the effectiveness of SHM

According to a former study[2], eliminating the NES domain of AID could increase the mutation rate of SHM by up to 40 folds compared to AID(wild type). Since the catalytic domain of CSR and NES domain overlap at the C-terminus 16 a.a, eliminating the C-terminus 16 a.a can increase the efficiency of SHM while inhibiting the function of CSR.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 76
    Illegal SapI site found at 177