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Volumn 275, Issue 5299, 1997, Pages 540-543

Requirement for the transcription factor LSIRF/IRF4 for mature B and T lymphocyte function

Author keywords

[No Author keywords available]

Indexed keywords

TRANSCRIPTION FACTOR;

EID: 15444346834     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5299.540     Document Type: Article
Times cited : (509)

References (31)
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    • Here we use the name IRF4, which was suggested and adopted for LSIRF by the nomenclature committee of the Genome Data Base for the gene locus in humans.
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    • To construct the targeting vector, an 8.4-kb Hind III fragment containing the promoter and exons 1 to 6 of the IRF4 gene was isolated from a 129J genomic library (1). The neomycin resistance cassette of pKJ1 (neo) [V. L. J. Tybulewicz et al., Cell 65, 1153 (1991)]; a Bam Hi-Hind III fragment containing IRF4 exons 4, 5, and 6 (long arm); and a polymerase chain reaction (PCR) fragment of intron 1 (short arm) were sequentially cloned into pBluescript SK. The targeting vector was linearized, and E14 embryonic stem (ES) cells were transfected and selected with G418. Targeted ES cell colonies were screened by PCR with primers specific for the 3′ region of neo and for a genomic sequence 5′ of the targeting construct. Positive ES cell lines were verified by Southern hybridization after Hind III digestion, with the use of probes specific for a sequence 5′ of the short arm (5′ probe) or for neo. Out of 1500 G418-resistant colonies, we could isolate 8 with the correct mutation and with a single neo integration. Positive ES cells were injected into CD1 blastocysts, and chimeric offspring were mated with C57BL/6 female mice. Mice were screened by Southern blot analysis or by PCR of tail DNA with the primers 5′-GCA ATG GGA AAC TCC GAC AGT-3′ and 5′-CAG CGT CCT CCT CAC GAT TGT-3′, specific for exon 2; and primers 5′-CCG GTG CCC TGA ATG AAC TGC-3′ and 5′-CAA TAT CAC GGG TAG CCA ACG-3′, specific for neo.
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    • note
    • Healthy 5-to 7-week-old mice were used for all experiments. Mice were maintained according to the guidelines of the Canadian Council of Animal Care. H.-W.M. is a recipient of a grant from the Deutsche Forschungsgemeinschaft. The authors thank M. Bachmann, G. Boehmelt, G. Duncan, D. Ferrick, C. Furlonger, D. Kägi, F. Kiefer, L. Marengere, C. Paige, J. Penninger, M. Saunders, R. Schmits, D. Siderovski, D. Speiser, S. Yoshinaga, and P. Waterhouse for providing reagents and for helpful comments.


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