Authors
Gothwal, S. K.
Abstract
Activation-Induced Cytidine Deaminase (AID) induces DNA double-strand breaks (DSBs) at the switch (S) regions of the Immunoglobulin heavy chain (Igh) locus, which are essential for Class Switch Recombination (CSR) and Somatic Hypermutation (SHM), key processes for effective antibody production. While AID activity is critical, its off-target effects, such as DSBs at the Myc locus, can cause chromosomal translocations like IghMyc fusions, contributing to B-cell lymphomas. The factors assembled on Igh locus that tether AID-induced DSBs and subsequently the CSR, remain unknown. To address this, we developed a method to isolate CSR-specific factors by inserting a 5X-GAL4UAS sequence at the switch mu (Smu) region in CH12 cells. This engineered site enables recruitment of a 3FLAGGAL4 DNA-binding protein (3F-GAL4DBD), allowing specific pulldown of proteins enriched at the Smu region. Successful recovery of BRD2 from the Smu region, a known CSR regulator, validated this approach. Characterizing these factors may uncover novel regulators of CSR and highlight mechanisms that balance antibody diversification with genomic integrity in B cells.
Preprint server:
bioRxiv
The authors list and abstract were imported from bioRxiv on 07 Nov 2025.
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