Wild-type V(D)J recombination in scid pre-B cells.
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-J kappa joints, as assayed by a polymerase chain reaction and DNA sequencing. In addition, three immunoglobulin mu- scid pre-B cell lines showed both scid and wild-type V-J kappa 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.
Hendrickson
E A
EA
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, Massachusetts.
Schlissel
M S
MS
Weaver
D T
DT
D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004254:DNA Nucleotidyltransferases; D015322:Gene Rearrangement, B-Lymphocyte; D005803:Genes, Immunoglobulin; D007145:Immunoglobulin kappa-Chains / Q000235:genetics*; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D007153:Immunologic Deficiency Syndromes / Q000235:genetics*; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D045567:VDJ Recombinases
D007145:Immunoglobulin kappa-Chains; D007148:Immunoglobulin mu-Chains; D009841:Oligonucleotides; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases