Liver-specific expression of a phosphoenolpyruvate carboxykinase-neo gene in genetically modified chickens.
In order to investigate the potential of the avian liver for the expression of recombinant proteins in vivo, replication-competent retroviral vectors were used to introduce a recombinant rat phosphoenolpyruvate carboxykinase promoter-driven neomycin resistance gene (PEPCKneo) into early Line 11 Leghorn embryos. After hatching, these birds possessed apparently intact PEPCKneo sequences in most tissues examined, however, the neo protein was expressed preferentially in the liver (up to .45% of total cellular protein). Therefore, the tissue specificity of the PEPCK promoter from the rat was retained in the chicken, although hormone responsiveness was not observed. Retroviral vectors used to transmit the genes were more stable during passage in either fibroblast cells or in the animal if the inserted genes were oriented in the same (sense) direction as the viral genome. After Geneticin drug selection in cultured cells, PEPCKneo mRNA was the predominant recombinant species observed on Northern blots, whereas embryos expressed mostly the RNA species originating in the retroviral long terminal repeats. The results demonstrate the potential usefulness of liver-specific gene expression in chickens, as well as the transcriptional effects observed when a foreign promoter is introduced into the replication-competent vector.
Laboratories of Molecular and Developmental Biology, Ohio State University, Wooster 44691.
R F
RF
Cook
S J
SJ
Cook
S
S
Savon
M
M
McGrane
M
M
Hartitz
R W
RW
Hanson
C P
CP
Hodgson
D000375:Aging / Q000502:physiology; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy; D019868:Kanamycin Kinase; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000096:biosynthesis / Q000235:genetics*; D010770:Phosphotransferases / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011784:Quail; D051381:Rats; D014162:Transfection
D010770:Phosphotransferases; D019868:Kanamycin Kinase; D010729:Phosphoenolpyruvate Carboxykinase (GTP)