Combination of 3' and 5' IgH regulatory elements mimics the B-specific endogenous expression pattern of IgH genes from pro-B cells to mature B cells in a transgenic mouse model.
To ensure the B cell differentiation stage specificity of the intronic Emu element and of the locus control region (LCR) that lies downstream of the IgH chain locus, we generated transgenic mice harboring a V(H) promoter-GFP reporter gene linked to the 3'LCR region and the Emu element. By flow cytometry, GFP(+) lymphocytes were observed amongst pro-B cells (B220(+)CD43(+)CD117(+)) and at all stages of differentiation up to mature B cells (B220(+)IgM(+)IgD(+)). Expression was strictly confined to cells committed to the B lymphocyte lineage as judged by the lack of GFP(+)Thy1,2(+) cells (T lymphocytes) and GFP(+)B220(-)CD117(+)CD43(+) cells (uncommitted lymphohematopoietic progenitors). Therefore, the Emu-GFP-3'LCR transgene is not expressed by hematopoietic stem cells, begins its expression in pro-B cells and is specifically active at all stages of B cell maturation. The combination of 3' and 5' IgH regulatory elements thus appears as a potentially useful cassette in transgenes that require a stringent and early B lineage-specific expression.
UMR CNRS 6101, Faculté de Médecine, 2 rue Dr Marcland, 87025, Limoges, France. laurence.gugliemi@unilim.fr
Laurence
L
Guglielmi
Marc
M
Le Bert
Isabelle
I
Comte
Marie Laure
ML
Dessain
Mireille
M
Drouet
Christiane
C
Ayer-Le Lievre
Michel
M
Cogné
Yves
Y
Denizot
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D019175:DNA Methylation; D005786:Gene Expression Regulation; D005803:Genes, Immunoglobulin / Q000235:genetics*; D016654:Genes, RAG-1 / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D013154:Spleen / Q000166:cytology; D019076:Transgenes / Q000235:genetics
D007143:Immunoglobulin Heavy Chains