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Related Concept Videos

Homologous Recombination02:31

Homologous Recombination

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...
Homologous Recombination02:31

Homologous Recombination

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...
Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
Exon Recombination02:32

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon has three reading...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Base-pairing and DNA Repair02:27

Base-pairing and DNA Repair

Erwin Chargaff’s rules on DNA equivalence paved the way for the discovery of base pairing in DNA. Chargaff’s rules state that in a double-stranded DNA molecule,

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Related Experiment Video

Updated: Jul 2, 2026

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization
13:55

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

Published on: February 3, 2013

A biochemically defined system for coding joint formation in V(D)J recombination.

Haihui Lu1, Noriko Shimazaki1, Prafulla Raval2

  • 1Norris Comprehensive Cancer Center, Room 5428, University of Southern California Keck School of Medicine, 1441 Eastlake Avenue, MC9176, Los Angeles, CA 90089, USA.

Molecular Cell
|August 30, 2008
PubMed
Summary
This summary is machine-generated.

The Artemis:DNA-PKcs complex processes hairpin DNA ends during V(D)J recombination. This reconstituted system clarifies antigen receptor gene assembly and identifies key enzymes involved in DNA repair.

More Related Videos

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
15:07

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma

Published on: December 28, 2015

Related Experiment Videos

Last Updated: Jul 2, 2026

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization
13:55

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

Published on: February 3, 2013

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
15:07

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma

Published on: December 28, 2015

Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • V(D)J recombination is a complex biological process essential for adaptive immunity.
  • The RAG complex initiates this process by creating double-stranded DNA breaks and hairpin coding ends.
  • Understanding the precise mechanisms of V(D)J recombination is crucial for comprehending immune system development.

Purpose of the Study:

  • To elucidate the role of the Artemis:DNA-PKcs complex in processing hairpin DNA ends during V(D)J recombination.
  • To reconstitute V(D)J recombination in vitro using purified components to study antigen receptor diversification.
  • To identify the enzymes responsible for end modification during V(D)J recombination.

Main Methods:

  • Activation of the Artemis:DNA-PKcs complex by hairpin DNA ends in cis.
  • Reconstitution of V(D)J recombination using 13 purified polypeptides.
  • Analysis of recombination site features including nucleolytic resection, P nucleotides, and N nucleotide addition.

Main Results:

  • The kinase activity of Artemis:DNA-PKcs is activated by hairpin DNA ends, leading to nicking.
  • A fully defined system using 13 polypeptides successfully reconstituted V(D)J recombination, adhering to the 12/23 rule.
  • The reconstituted system generated recombination sites with features identical to those observed in vivo.

Conclusions:

  • The Artemis:DNA-PKcs complex plays a critical role in nicking hairpin DNA ends during V(D)J recombination.
  • Most, if not all, enzymes involved in V(D)J recombination end modification have been identified.
  • This study provides a comprehensive in vitro system for studying antigen receptor gene assembly.