High-level and erythroid-specific expression of human glucose-6-phosphate dehydrogenase in transgenic mice.
Human Glc-6-P dehydrogenase (Glc-6-P) cDNA spanning the entire coding region was subcloned into a pSG5 vector that contains an early SV40 promoter, intron II of the rabbit beta-globin gene, and a polyadenylation signal. This expression cassette was then placed downstream of the human beta-globin locus control region and injected into fertilized mouse eggs. Among five transgenic founders that contained intact copies of the construct, one founder expressed human Glc-6-P dehydrogenase enzyme in a high-level and erythroid-specific fashion (5 x higher than endogenous Glc-6-P dehydrogenase activity). When this male founder mated with a normal individual, all the offspring that carried the transgene showed high-level expression of Glc-6-P dehydrogenase activity in erythroid cells. The endogenous mouse Glc-6-P dehydrogenase in all high-expression mice could be competed out by forming a hybrid with human Glc-6-P dehydrogenase. Our results indicate that the locus control region can drive the human Glc-6-P dehydrogenase gene to be specifically expressed in the erythroid cells of transgenic mice. The results described here provide a basis for experiments designed to express human Glc-6-P dehydrogenase in transgenic mice and suggest a suitable approach to producing a mouse model for studying human Glc-6-P dehydrogenase deficiency.
Tang
T K
TK
Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, Republic of China.
Tam
K B
KB
Huang
S C
SC
D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004912:Erythrocytes / Q000201:enzymology*; D005260:Female; D005822:Genetic Vectors; D005914:Globins / Q000235:genetics; D005954:Glucosephosphate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics*; D006461:Hemolysis; D006801:Humans; D007700:Kinetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010375:Pedigree; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011817:Rabbits; D013539:Simian virus 40 / Q000235:genetics
D005914:Globins; D004247:DNA; D005954:Glucosephosphate Dehydrogenase