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Related Concept Videos

Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Related Experiment Video

Updated: Dec 13, 2025

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CTCF orchestrates long-range cohesin-driven V(D)J recombinational scanning.

Zhaoqing Ba1,2, Jiangman Lou3,4, Adam Yongxin Ye3,4

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CTCF binding impedes RAG scanning during V(D)J recombination. Removing CTCF allows RAG to access distal V(D)S segments, promoting efficient V(D)J recombination in progenitor B cells.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • The RAG endonuclease initiates V(D)J recombination in progenitor B cells, a crucial process for adaptive immunity.
  • RAG scanning, potentially aided by cohesin, locates DNA segments for recombination, but CTCF-bound elements can impede this process.
  • Distal V(H) segment utilization is thought to rely on locus contraction and diffusional access to the recombination center.

Purpose of the Study:

  • To investigate the role of linear RAG scanning in distal V(H) segment usage.
  • To determine the impact of cohesin and CTCF on RAG scanning and V(H)-to-DJ(H) joining in G1-arrested pro-B cells.
  • To explore how modulating CTCF activity affects Igh locus accessibility and recombination.

Main Methods:

  • Utilized an auxin-inducible system to degrade RAD21 (a cohesin component) or CTCF in G1-arrested v-Abl pro-B cell lines.
  • Assessed V(D)J recombination efficiency and chromatin interactions following protein degradation.
  • Compared recombination patterns in manipulated cells with those of 'locus-contracted' primary pro-B cells.

Main Results:

  • RAD21 degradation abolished V(D)J recombination and RAG scanning interactions, except for diffusion-mediated DQ52-to-J(H) joining.
  • CTCF degradation suppressed CTCF-bound element interactions but significantly promoted distal V(H) segment recombination and interactions.
  • The recombination patterns after CTCF degradation mimicked those observed in locus-contracted primary pro-B cells.

Conclusions:

  • Cohesin-mediated RAG scanning is essential for V(D)J recombination.
  • CTCF acts as a barrier to RAG scanning, preventing access to distal V(H) segments.
  • Downregulating CTCF facilitates cohesin-driven RAG scanning across the Igh locus, promoting distal V(H) utilization.