The 12/23 rule is enforced at the cleavage step of V(D)J recombination in vivo

Background: V(D)J recombination is initiated by the introduction of double‐stranded breaks (DSB) adjacent to recombination signal sequences (RSS). Each RSS contains a conserved heptamer and a conserved nonamer element separated by a 12 or 23 nucleotide spacer. In vivo, efficient recombination requires one RSS of each spacer length, although it has been unclear whether this ‘12/23 rule’ regulates cleavage, joining, or both.

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