V(D)J recombination: In vitro coding joint formation

Frances Weis-Garcia, Eva Besmer, Dennis J. Sawchuk, Wong Yu, Yun Hu, Sylvanie Cassard, Michel C. Nussenzweig, Patricia Cortes

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Antigen receptor genes are assembled through a mechanism known as V(D)J recombination, which involves two different joining reactions: signal and coding joining. Formation of these joints is essential for antigen receptor assembly as well as maintaining chromosomal integrity. Here we report on a cell-free system for coding joint formation using deletion and inversion recombination substrates. In vitro coding joint formation requires RAG1, RAG2, and heat-labile factors present in the nuclear extract of nonlymphoid cells. Both inversion- and deletion-mediated coding joint reactions produce diverse coding joints, with deletions and P nucleotide addition. We also show that deletion-mediated coding joint formation follows the 12/23 rule and requires the catalytic subunit of DNA-dependent protein kinase.

Original languageEnglish
Pages (from-to)6379-6385
Number of pages7
JournalMolecular and Cellular Biology
Volume17
Issue number11
DOIs
StatePublished - Nov 1997
Externally publishedYes

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