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V(D)J recombination is not activated by demethylation of the kappa locus

Cherry, Sara R. and Beard, Caroline and Jaenisch, Rudolf and Baltimore, David (2000) V(D)J recombination is not activated by demethylation of the kappa locus. Proceedings of the National Academy of Sciences of the United States of America, 97 (15). pp. 8467-8472. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:CHEpnas00

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Abstract

V(D)J recombination is thought to be regulated by changes in the accessibility of target sites, such as modulation of methylation. To test whether demethylation of the kappa locus can activate recombination, we generated two recombinationally active B cell lines in which the gene for maintenance of genomic DNA methylation, Dnmt1, was flanked with loxP sites. Transduction with a retrovirus expressing both the cre recombinase and green fluorescent protein allowed us to purify recombinationally active cells devoid of methylation. Loss of methylation of the kappa locus was not sufficient to activate recombination, although transcription was activated in one line. It appears that demethylation of the kappa locus is not the rate-limiting step for altering accessibility and thus regulated demethylation does not generate specificity of recombination.


Item Type:Article
Additional Information:© 2000 by The National Academy of Sciences. Contributed by David Baltimore, May 15, 2000. We thank members of the Jaenisch and Baltimore labs for advice and discussions. We are grateful to Dr. Luk van Parijs for advice on retroviral infections; Dr. Christopher Roman for assistance in generating Abelson lines and for discussions and comments on the manuscript; and Glen Paradis for fluorescence-activated cell sorting and technical assistance. D.B. is supported by a National Institutes of Health grant.
Subject Keywords:REGION GENE SEGMENTS, IMMUNOGLOBULIN GENES, B-CELLS, METHYLATION STATUS, LYMPHOID-CELLS, HELPER-FREE, HIGH-TITER, IN-VIVO, TRANSCRIPTION, REARRANGEMENT
Record Number:CaltechAUTHORS:CHEpnas00
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:CHEpnas00
Alternative URL:http://dx.doi.org/10.1073/pnas.150218497
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:575
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:25 Aug 2005
Last Modified:14 Nov 2014 19:18

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