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 Legh...

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Bibliographic Details
Published in:Poultry science 1993-03, Vol.72 (3), p.554-567
Main Authors: Cook, R.F. (University of Kentucky, Lexington, KY), Cook, S.J, Savon, S, McGrane, M, Hartitz, M, Hanson, R.W, Hodgson, C.P
Format: Article
Language:English
Subjects:
Aging - physiology
ANIMAL TRANSGENIQUE
ANIMALES TRANSGENICOS
Animals
Animals, Genetically Modified
Cell Line
Chick Embryo
Chickens
Enzyme-Linked Immunosorbent Assay
EXPRESION GENICA
EXPRESSION DES GENES
FOIE
Genetic Therapy
GENIE GENETIQUE
HIBRIDACION DE ADN
HIGADO
HYBRIDATION D'ADN
INGENIERIA GENETICA
Kanamycin Kinase
Kidney - enzymology
Liver - enzymology
MARCADORES GENETICOS
MARQUEUR GENETIQUE
Myocardium - enzymology
Organ Specificity
Phosphoenolpyruvate Carboxykinase (GTP) - biosynthesis
Phosphoenolpyruvate Carboxykinase (GTP) - genetics
Phosphotransferases - biosynthesis
Phosphotransferases - genetics
POLLO
POULET
Promoter Regions, Genetic
Quail
Rats
RETROVIRIDAE
Transfection
VECTEUR DE MALADIE
VECTORES
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Summary: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 0.45% 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
ISSN:0032-5791
1525-3171
DOI:10.3382/ps.0720554