Mechanisms That Control Antigen Receptor Variable Region Exon Assembly

Summary

This research project aims to elucidate the distinct mechanisms by which Ig heavy chain (Igh) and Ig light chain (Igk) loci achieve long-range V(D)J recombination, a process critical for generating antibody diversity.

What they want

The project proposes two specific aims to compare and contrast the long-range V(D)J recombination mechanisms of Igh and Igk. Aim 1 will test the hypothesis that Igh uses linear RAG chromatin scanning leading to predominantly deletional recombination. Aim 2 will investigate if Igk is structurally optimized for a related loop extrusion-based mechanism accommodating both deletional and inverted Vk-to-Jk joining. Studies will utilize advanced technologies developed by the researchers, including LAM-HTGTS-V(D)J-Seq and LAM-3C-HTGTS, and G1-arrested, RAG inducible v-Abl transformed pro-B cell approaches, with key results confirmed in normal progenitor and precursor B cells.

Deliverables

New paradigms for understanding V(D)J recombination in vivo
Major new insights into fundamental mechanisms that establish highly diverse primary antibody repertoires
Critical information for understanding how RAG targets cryptic RSSs in other genes
Understanding of how RAG promotes common translocations or interstitial deletions in B and T lymphocyte cancers

Technical requirements

LAM-HTGTS-V(D)J-Seq technology
LAM-3C-HTGTS technology
G1-arrested, RAG inducible v-Abl transformed pro-B cell cell ("v-Abl cell") approaches
Introduced Igh or Igk locus modifications
Targeted protein depletion