Wild-type V(D)J recombination in scid Pre-B cells

Eric A. Hendrickson, Mark S. Schlissel, David T. Weaver

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Homozygous mutation at the scid locus in the mouse results in the aberrant rearrangement of immunoglobulin and T-cell receptor gene segments. We introduced a retroviral vector containing an inversional immunoglobulin rearrangement cassette into scid pre-B cells. Most rearrangements were accompanied by large deletions, consistent with previously characterized effects of the scid mutation. However, two cell clones were identified which contained perfect reciprocal fragments and wild-type coding joints, documenting, on a molecular level, the ability of scid pre-B cells to generate functional protein-coding domains. Subsequent rearrangement of the DGR cassette in one of these clones was accompanied by a deletion, suggesting that this cell clone had not reverted the scid mutation. Indeed, induced rearrangement of the endogenous kappa loci in these two cell clones resulted in a mixture of scid and wild-type V-Jκ joints, as assayed by a polymerase chain reaction and DNA sequencing. In addition, three immunoglobulin μ- scid pre-B cell lines showed both scid and wild-type V-Jκ joins. These experiments strongly suggest that the V(D)J recombinase activity in scid lymphoid cells is diminished but not absent, consistent with the known leakiness of the scid mutation.

Original languageEnglish (US)
Pages (from-to)5397-5407
Number of pages11
JournalMolecular and cellular biology
Volume10
Issue number10
DOIs
StatePublished - 1990

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