regulation of rearrangement; mouse immunoglobulin genes; allelic exclusion; germline promoter; positive regulatory element
Abstract :
[en] We have shown previously that a mutation of the KI-KII site immediately 5' to J(kappa)1 on the mouse immunoglobulin light chain kappa locus reduces the rearrangement level in cis, although it does not affect transcription. Here we deleted by homologous recombination in mouse embryonic stem cells a 4-kb DNA fragment, located immediately upstream of the KI-KII element, which contains the promoter of the long germline transcript. Analysis of gene-targeted heterozygous mouse splenic B cells showed a strong decrease in rearrangement for the allele bearing the deletion. When both the KI-KII mutation and the 4-kb deletion were present on the same allele, the overall reduction in rearrangement was stronger than with the 4-kb deletion alone underlying the role of these two elements in the regulation of rearrangement. The same deletion was performed by homologous recombination on one allele of the rearrangement-inducible mouse 103/bcl2-hygro(R) pre-B cell line, and resulted in a similar reduction in the induction of rearrangement of the mutated allele. This result validates this cell line as an in vitro model for studying the incidence of gene-targeted modifications of the kappa locus on the regulation of rearrangement
Cocea, L.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
De Smet, A.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
Saghatchian, M.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
Fillatreau, S.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
Ferradini, L.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
Schurmans, Stéphane ; Université Libre de Bruxelles - ULB > Institut de Recherche Interdisciplinaire en Biologie Humaine et Nucléaire, Institut de Biologie et de Médecine Moléculaire
Weill, J. C.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
Reynaud, A.; Institut National de la Santé et de la Recherche Médicale, Unité 373, Faculté de Médecine Necker, Paris, France
Language :
English
Title :
A targeted deletion of a region upstream from the Jkappa cluster impairs kappa chain rearrangement in cis in mice and in the 103/bcl2 cell line
Publication date :
1999
Journal title :
Journal of Experimental Medicine
ISSN :
0022-1007
eISSN :
1540-9538
Publisher :
Rockefeller University Press, New York, United States - New York
Volume :
189
Pages :
1443-1450
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Abbreviations used in this paper: BCR, B cell receptor; ES, embryonic stem; RT, reverse transcription
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