Genetic disruption of the murine complement C3 promoter region generates deficient mice with extrahepatic expression of C3 mRNA.
Genetic deficiencies of the complement protein C3 occur naturally in humans and animal models and have been induced in mice by targeted deletion of the C3 gene. The study of these deficiencies has provided evidence for C3 functions in immune responses. C3 deficient mice were generated by replacing the 5'-flanking region of the C3 gene with the neomycin-resistance (neo) gene. Serum from these mice had no detectable C3 protein or complement activity. Challenge with Streptococcus pneumoniae revealed approximately 2000-fold increase in bacteremia as compared to littermate controls. C3 mRNA was absent in the liver, but it was detected in the lung, kidney, fat tissue, heart and spleen. Metabolic labeling of the lung tissue and peritoneal macrophages showed synthesis of pro-C3, but no post-synthetic intracellular processing of the protein and no secretion of mature C3. cDNA analysis at the cap site indicated that extrahepatic transcription of the targeted gene was initiated in the neo cassette, close to the C3/neo junction and predicted a primary translation product lacking the leader peptide. The data indicate that these mice provide a good animal model for the study of complete C3 deficiencies and a potential probe for tissue-specific C3 gene regulatory elements.
Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA.
A
A
Circolo
G
G
Garnier
W
W
Fukuda
X
X
Wang
T
T
Hidvegi
A J
AJ
Szalai
D E
DE
Briles
J E
JE
Volanakis
R A
RA
Wetsel
H R
HR
Colten
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D003176:Complement C3 / Q000096:biosynthesis / Q000172:deficiency* / Q000235:genetics* / Q000276:immunology; D004247:DNA / Q000235:genetics; D005091:Exons; D005260:Female; D017353:Gene Deletion; D015870:Gene Expression; D018390:Gene Targeting; D007113:Immunity, Innate; D017403:In Situ Hybridization; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011008:Pneumococcal Infections / Q000276:immunology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013296:Streptococcus pneumoniae
D003176:Complement C3; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase