Retroviral-mediated gene transfer of human phenylalanine hydroxylase into NIH 3T3 and hepatoma cells

Journal Article · Wed Jan 01 00:00:00 EST 1986 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.83.2.409· OSTI ID:6220283

Ledley, F D; Grenett, H E; McGinnis-Shelnutt, M; Woo, S L.C.

Phenylketonuria (PKU) is caused by deficiency of the hepatic enzyme phenylalanine hydroxylase (PAH). A full-length human PAH cDNA sequence has been inserted into pzip-neoSV(X), which is a retroviral vector containing the bacterial neo gene. The recombinant has been transfected into Psi2 cells, which provide synthesis of the retroviral capsid. Recombinant virus was detected in the culture medium of the transfected Psi2 cells, which is capable of transmitting the human PAH gene into mouse NIH 3T3 cells by infection leading to stable incorporation of the recombinant provirus. Infected cells express PAH mRNA, immunoreactive PAH protein, and exhibit pterin-dependent phenylaline hydroxylase activity. The recombinant virus is also capable of infecting a mouse hepatoma cell line that does not normal synthesize PAH. PAH activity is present in the cellular extracts and the entire hydroxylation system is reconstituted in the hepatoma cells infected with the recombinant viruses. Thus, recombinant viruses containing human PAH cDNA provide a means for introducing functional PAH into mammalian cells of hepatic origin and can potentially be introduced into whole animals as a model for somatic gene therapy for PKU.

• Research Organization: Baylor College of Medicine, Houston, TX
• OSTI ID: 6220283
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:2
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
HYDROXYLASES
ENZYME ACTIVITY
LIVER CELLS
GENETIC ENGINEERING
TUMOR CELLS
METABOLIC DISEASES
PHENYLALANINE
PLASMIDS
RECOMBINANT DNA
VIRUSES
AMINO ACIDS
ANIMAL CELLS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
DISEASES
DNA
ENZYMES
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PARASITES
SOMATIC CELLS
550400* - Genetics