Part:BBa_K4151016

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.

References

Zan H, Casali P. Regulation of AICDA expression and AID activity. Autoimmunity. 2013 Mar;46(2):83-101. doi: 10.3109/08916934.2012.749244. Epub 2013 Jan 17. PMID: 23181381; PMCID: PMC3762583.)

Ito S, Nagaoka H, Shinkura R, Begum N, Muramatsu M, Nakata M, Honjo T. Activation-induced cytidine deaminase shuttles between nucleus and cytoplasm like apolipoprotein B mRNA editing catalytic polypeptide 1. Proc Natl Acad Sci U S A. 2004 Feb 17;101(7):1975-80. doi: 10.1073/pnas.0307335101. Epub 2004 Feb 9. PMID: 14769937; PMCID: PMC357037.

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