Retroviral-Mediated Gene Transfer and Expression of Human Phenylalanine Hydroxylase in Primary Mouse Hepatocytes
Genetic therapy for phenylketonuria (severe phenylalanine hydroxylase deficiency) may require introduction of a normal phenylalanine hydroxylase gene into hepatic cells of patients. We report development of a recombinant retrovirus based on the N2 vector for gene transfer and expression of human phe...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1988-11, Vol.85 (21), p.8146-8150 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Peng, Hong Armentano, Donna MacKenzie-Graham, Leslie Shen, Rong-Fong Darlington, Gretchen Ledley, Fred D. Savio L. C. Woo |
| description | Genetic therapy for phenylketonuria (severe phenylalanine hydroxylase deficiency) may require introduction of a normal phenylalanine hydroxylase gene into hepatic cells of patients. We report development of a recombinant retrovirus based on the N2 vector for gene transfer and expression of human phenylalanine hydroxylase cDNA in primary mouse hepatocytes. This construct contains an internal promoter of the human α 1-antitrypsin gene driving transcription of the phenylalanine hydroxylase cDNA. Primary mouse hepatocytes were isolated from newborn mice, infected with the recombinant virus, and selected for expression of the neomycin-resistance gene. Hepatocytes transformed with the recombinant virus contained high levels of human phenylalanine hydroxylase mRNA transcripts originating from the retroviral and internal promoters. These results demonstrate that the transcriptional regulatory elements of the α 1-antitrypsin gene retain their tissue-specific function in the recombinant provirus and establish a method for efficient transfer and high-level expression of human phenylalanine hydroxylase in primary hepatocytes. |
| doi_str_mv | 10.1073/pnas.85.21.8146 |
| format | Article |
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C. Woo</creator><creatorcontrib>Peng, Hong ; Armentano, Donna ; MacKenzie-Graham, Leslie ; Shen, Rong-Fong ; Darlington, Gretchen ; Ledley, Fred D. ; Savio L. C. Woo</creatorcontrib><description>Genetic therapy for phenylketonuria (severe phenylalanine hydroxylase deficiency) may require introduction of a normal phenylalanine hydroxylase gene into hepatic cells of patients. We report development of a recombinant retrovirus based on the N2 vector for gene transfer and expression of human phenylalanine hydroxylase cDNA in primary mouse hepatocytes. This construct contains an internal promoter of the human α 1-antitrypsin gene driving transcription of the phenylalanine hydroxylase cDNA. Primary mouse hepatocytes were isolated from newborn mice, infected with the recombinant virus, and selected for expression of the neomycin-resistance gene. Hepatocytes transformed with the recombinant virus contained high levels of human phenylalanine hydroxylase mRNA transcripts originating from the retroviral and internal promoters. These results demonstrate that the transcriptional regulatory elements of the α 1-antitrypsin gene retain their tissue-specific function in the recombinant provirus and establish a method for efficient transfer and high-level expression of human phenylalanine hydroxylase in primary hepatocytes.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.85.21.8146</identifier><identifier>PMID: 3186716</identifier><identifier>CODEN: PNASA6</identifier><language>eng</language><publisher>Washington, DC: National Academy of Sciences of the United States of America</publisher><subject>550401 - Genetics- Tracer Techniques ; AMINO ACIDS ; ANIMAL CELLS ; ANIMALS ; BASIC BIOLOGICAL SCIENCES ; BETA DECAY RADIOISOTOPES ; BETA-MINUS DECAY RADIOISOTOPES ; Biological and medical sciences ; Biotechnology ; Blotting, Southern ; CARBOXYLIC ACIDS ; Cell lines ; CELL TRANSFORMATIONS ; Complementary DNA ; DAYS LIVING RADIOISOTOPES ; DISEASES ; DNA ; DNA - analysis ; ENZYMES ; Fundamental and applied biological sciences. Psychology ; Gene expression ; Gene Expression Regulation ; GENE RECOMBINATION ; GENE REGULATION ; GENE REPRESSORS ; Gene therapy ; Genes ; Genetic engineering ; Genetic technics ; Health. Pharmaceutical industry ; Hepatocytes ; HEREDITARY DISEASES ; Humans ; HYDROXYLASES ; Industrial applications and implications. Economical aspects ; ISOTOPES ; LIGHT NUCLEI ; Liver ; Liver - enzymology ; LIVER CELLS ; MAMMALS ; MESSENGER-RNA ; Methods. Procedures. Technologies ; MICE ; MICROORGANISMS ; Molecular and cellular biology ; Molecular genetics ; NUCLEI ; NUCLEIC ACIDS ; NUCLEOPROTEINS ; ODD-ODD NUCLEI ; ORGANIC ACIDS ; ORGANIC COMPOUNDS ; OXIDOREDUCTASES ; PARASITES ; PHENYLALANINE ; Phenylalanine Hydroxylase - genetics ; Phenylketonuria ; PHOSPHORUS 32 ; PHOSPHORUS ISOTOPES ; PROTEINS ; RADIOISOTOPES ; RECOMBINANT DNA ; Recombination, Genetic ; Retroviridae ; Retroviridae - genetics ; RNA ; RNA, Messenger - metabolism ; RODENTS ; SOMATIC CELLS ; THERAPY ; TRANSCRIPTION ; Transcription, Genetic ; Transfection ; VERTEBRATES ; VIRUSES</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1988-11, Vol.85 (21), p.8146-8150</ispartof><rights>1990 INIST-CNRS</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c559t-75a3e6b0000a9005b80a80f8b41ac656f59a52ecacf103d975213437da5300433</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/85/21.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/32587$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/32587$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,777,781,800,882,27905,27906,57998,58231</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6837754$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7229634$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3186716$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/5546707$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Peng, Hong</creatorcontrib><creatorcontrib>Armentano, Donna</creatorcontrib><creatorcontrib>MacKenzie-Graham, Leslie</creatorcontrib><creatorcontrib>Shen, Rong-Fong</creatorcontrib><creatorcontrib>Darlington, Gretchen</creatorcontrib><creatorcontrib>Ledley, Fred D.</creatorcontrib><creatorcontrib>Savio L. C. Woo</creatorcontrib><title>Retroviral-Mediated Gene Transfer and Expression of Human Phenylalanine Hydroxylase in Primary Mouse Hepatocytes</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Genetic therapy for phenylketonuria (severe phenylalanine hydroxylase deficiency) may require introduction of a normal phenylalanine hydroxylase gene into hepatic cells of patients. We report development of a recombinant retrovirus based on the N2 vector for gene transfer and expression of human phenylalanine hydroxylase cDNA in primary mouse hepatocytes. This construct contains an internal promoter of the human α 1-antitrypsin gene driving transcription of the phenylalanine hydroxylase cDNA. Primary mouse hepatocytes were isolated from newborn mice, infected with the recombinant virus, and selected for expression of the neomycin-resistance gene. Hepatocytes transformed with the recombinant virus contained high levels of human phenylalanine hydroxylase mRNA transcripts originating from the retroviral and internal promoters. These results demonstrate that the transcriptional regulatory elements of the α 1-antitrypsin gene retain their tissue-specific function in the recombinant provirus and establish a method for efficient transfer and high-level expression of human phenylalanine hydroxylase in primary hepatocytes.</description><subject>550401 - Genetics- Tracer Techniques</subject><subject>AMINO ACIDS</subject><subject>ANIMAL CELLS</subject><subject>ANIMALS</subject><subject>BASIC BIOLOGICAL SCIENCES</subject><subject>BETA DECAY RADIOISOTOPES</subject><subject>BETA-MINUS DECAY RADIOISOTOPES</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Blotting, Southern</subject><subject>CARBOXYLIC ACIDS</subject><subject>Cell lines</subject><subject>CELL TRANSFORMATIONS</subject><subject>Complementary DNA</subject><subject>DAYS LIVING RADIOISOTOPES</subject><subject>DISEASES</subject><subject>DNA</subject><subject>DNA - analysis</subject><subject>ENZYMES</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>GENE RECOMBINATION</subject><subject>GENE REGULATION</subject><subject>GENE REPRESSORS</subject><subject>Gene therapy</subject><subject>Genes</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Health. Pharmaceutical industry</subject><subject>Hepatocytes</subject><subject>HEREDITARY DISEASES</subject><subject>Humans</subject><subject>HYDROXYLASES</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>ISOTOPES</subject><subject>LIGHT NUCLEI</subject><subject>Liver</subject><subject>Liver - enzymology</subject><subject>LIVER CELLS</subject><subject>MAMMALS</subject><subject>MESSENGER-RNA</subject><subject>Methods. Procedures. Technologies</subject><subject>MICE</subject><subject>MICROORGANISMS</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>NUCLEI</subject><subject>NUCLEIC ACIDS</subject><subject>NUCLEOPROTEINS</subject><subject>ODD-ODD NUCLEI</subject><subject>ORGANIC ACIDS</subject><subject>ORGANIC COMPOUNDS</subject><subject>OXIDOREDUCTASES</subject><subject>PARASITES</subject><subject>PHENYLALANINE</subject><subject>Phenylalanine Hydroxylase - genetics</subject><subject>Phenylketonuria</subject><subject>PHOSPHORUS 32</subject><subject>PHOSPHORUS ISOTOPES</subject><subject>PROTEINS</subject><subject>RADIOISOTOPES</subject><subject>RECOMBINANT DNA</subject><subject>Recombination, Genetic</subject><subject>Retroviridae</subject><subject>Retroviridae - genetics</subject><subject>RNA</subject><subject>RNA, Messenger - metabolism</subject><subject>RODENTS</subject><subject>SOMATIC CELLS</subject><subject>THERAPY</subject><subject>TRANSCRIPTION</subject><subject>Transcription, Genetic</subject><subject>Transfection</subject><subject>VERTEBRATES</subject><subject>VIRUSES</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkd1rFDEUxYModV19FgRlkKJPs00mn_Mope0KLYrU55DN3KEps5MxyZTd_94MM1Z8sU-X3PPL5dx7EHpL8IZgSc-G3sSN4puKbBRh4hlaEVyTUrAaP0crjCtZKlaxl-hVjPcY45orfIJOKFFCErFCww9IwT-4YLryBhpnEjTFFfRQ3AbTxxZCYfqmuDgMAWJ0vi98W2zHvemL73fQHzvTmd5lfHtsgj_kd4TCZTG4vQnH4saPubGFwSRvjwnia_SiNV2EN0tdo5-XF7fn2_L629XX8y_XpeW8TqXkhoLYZcfY1BjzncJG4VbtGDFWcNHy2vAKrLEtwbSpJa8IZVQ2hlOMGaVr9HGe62NyOlqXwN5Z3_dgk-acCZnPt0afZmgI_tcIMem9ixa6vBNk41oqToSU_EmQ8EphplgGz2bQBh9jgFYP8yU0wXpKTE-JacV1RfSUWP7xfhk97vbQPPJLRFk_XXQTrenaHIt18RGTVVULyp7ChKJ5jwn7sGCTjT_qP3Y-_xfQ7dh1CQ4pk-9m8j4mH_76rriS9DehesvM</recordid><startdate>19881101</startdate><enddate>19881101</enddate><creator>Peng, Hong</creator><creator>Armentano, Donna</creator><creator>MacKenzie-Graham, Leslie</creator><creator>Shen, Rong-Fong</creator><creator>Darlington, Gretchen</creator><creator>Ledley, Fred D.</creator><creator>Savio L. C. Woo</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>19881101</creationdate><title>Retroviral-Mediated Gene Transfer and Expression of Human Phenylalanine Hydroxylase in Primary Mouse Hepatocytes</title><author>Peng, Hong ; Armentano, Donna ; MacKenzie-Graham, Leslie ; Shen, Rong-Fong ; Darlington, Gretchen ; Ledley, Fred D. ; Savio L. C. Woo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c559t-75a3e6b0000a9005b80a80f8b41ac656f59a52ecacf103d975213437da5300433</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>550401 - Genetics- Tracer Techniques</topic><topic>AMINO ACIDS</topic><topic>ANIMAL CELLS</topic><topic>ANIMALS</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>BETA DECAY RADIOISOTOPES</topic><topic>BETA-MINUS DECAY RADIOISOTOPES</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Blotting, Southern</topic><topic>CARBOXYLIC ACIDS</topic><topic>Cell lines</topic><topic>CELL TRANSFORMATIONS</topic><topic>Complementary DNA</topic><topic>DAYS LIVING RADIOISOTOPES</topic><topic>DISEASES</topic><topic>DNA</topic><topic>DNA - analysis</topic><topic>ENZYMES</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>GENE RECOMBINATION</topic><topic>GENE REGULATION</topic><topic>GENE REPRESSORS</topic><topic>Gene therapy</topic><topic>Genes</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Health. Pharmaceutical industry</topic><topic>Hepatocytes</topic><topic>HEREDITARY DISEASES</topic><topic>Humans</topic><topic>HYDROXYLASES</topic><topic>Industrial applications and implications. Economical aspects</topic><topic>ISOTOPES</topic><topic>LIGHT NUCLEI</topic><topic>Liver</topic><topic>Liver - enzymology</topic><topic>LIVER CELLS</topic><topic>MAMMALS</topic><topic>MESSENGER-RNA</topic><topic>Methods. Procedures. Technologies</topic><topic>MICE</topic><topic>MICROORGANISMS</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>NUCLEI</topic><topic>NUCLEIC ACIDS</topic><topic>NUCLEOPROTEINS</topic><topic>ODD-ODD NUCLEI</topic><topic>ORGANIC ACIDS</topic><topic>ORGANIC COMPOUNDS</topic><topic>OXIDOREDUCTASES</topic><topic>PARASITES</topic><topic>PHENYLALANINE</topic><topic>Phenylalanine Hydroxylase - genetics</topic><topic>Phenylketonuria</topic><topic>PHOSPHORUS 32</topic><topic>PHOSPHORUS ISOTOPES</topic><topic>PROTEINS</topic><topic>RADIOISOTOPES</topic><topic>RECOMBINANT DNA</topic><topic>Recombination, Genetic</topic><topic>Retroviridae</topic><topic>Retroviridae - genetics</topic><topic>RNA</topic><topic>RNA, Messenger - metabolism</topic><topic>RODENTS</topic><topic>SOMATIC CELLS</topic><topic>THERAPY</topic><topic>TRANSCRIPTION</topic><topic>Transcription, Genetic</topic><topic>Transfection</topic><topic>VERTEBRATES</topic><topic>VIRUSES</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Peng, Hong</creatorcontrib><creatorcontrib>Armentano, Donna</creatorcontrib><creatorcontrib>MacKenzie-Graham, Leslie</creatorcontrib><creatorcontrib>Shen, Rong-Fong</creatorcontrib><creatorcontrib>Darlington, Gretchen</creatorcontrib><creatorcontrib>Ledley, Fred D.</creatorcontrib><creatorcontrib>Savio L. C. Woo</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Peng, Hong</au><au>Armentano, Donna</au><au>MacKenzie-Graham, Leslie</au><au>Shen, Rong-Fong</au><au>Darlington, Gretchen</au><au>Ledley, Fred D.</au><au>Savio L. C. Woo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Retroviral-Mediated Gene Transfer and Expression of Human Phenylalanine Hydroxylase in Primary Mouse Hepatocytes</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1988-11-01</date><risdate>1988</risdate><volume>85</volume><issue>21</issue><spage>8146</spage><epage>8150</epage><pages>8146-8150</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><coden>PNASA6</coden><abstract>Genetic therapy for phenylketonuria (severe phenylalanine hydroxylase deficiency) may require introduction of a normal phenylalanine hydroxylase gene into hepatic cells of patients. We report development of a recombinant retrovirus based on the N2 vector for gene transfer and expression of human phenylalanine hydroxylase cDNA in primary mouse hepatocytes. This construct contains an internal promoter of the human α 1-antitrypsin gene driving transcription of the phenylalanine hydroxylase cDNA. Primary mouse hepatocytes were isolated from newborn mice, infected with the recombinant virus, and selected for expression of the neomycin-resistance gene. Hepatocytes transformed with the recombinant virus contained high levels of human phenylalanine hydroxylase mRNA transcripts originating from the retroviral and internal promoters. These results demonstrate that the transcriptional regulatory elements of the α 1-antitrypsin gene retain their tissue-specific function in the recombinant provirus and establish a method for efficient transfer and high-level expression of human phenylalanine hydroxylase in primary hepatocytes.</abstract><cop>Washington, DC</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>3186716</pmid><doi>10.1073/pnas.85.21.8146</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1988-11, Vol.85 (21), p.8146-8150 |
| issn | 0027-8424 1091-6490 |
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| recordid | cdi_proquest_miscellaneous_15280484 |
| source | MEDLINE; Jstor Complete Legacy; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | 550401 - Genetics- Tracer Techniques AMINO ACIDS ANIMAL CELLS ANIMALS BASIC BIOLOGICAL SCIENCES BETA DECAY RADIOISOTOPES BETA-MINUS DECAY RADIOISOTOPES Biological and medical sciences Biotechnology Blotting, Southern CARBOXYLIC ACIDS Cell lines CELL TRANSFORMATIONS Complementary DNA DAYS LIVING RADIOISOTOPES DISEASES DNA DNA - analysis ENZYMES Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Regulation GENE RECOMBINATION GENE REGULATION GENE REPRESSORS Gene therapy Genes Genetic engineering Genetic technics Health. Pharmaceutical industry Hepatocytes HEREDITARY DISEASES Humans HYDROXYLASES Industrial applications and implications. Economical aspects ISOTOPES LIGHT NUCLEI Liver Liver - enzymology LIVER CELLS MAMMALS MESSENGER-RNA Methods. Procedures. Technologies MICE MICROORGANISMS Molecular and cellular biology Molecular genetics NUCLEI NUCLEIC ACIDS NUCLEOPROTEINS ODD-ODD NUCLEI ORGANIC ACIDS ORGANIC COMPOUNDS OXIDOREDUCTASES PARASITES PHENYLALANINE Phenylalanine Hydroxylase - genetics Phenylketonuria PHOSPHORUS 32 PHOSPHORUS ISOTOPES PROTEINS RADIOISOTOPES RECOMBINANT DNA Recombination, Genetic Retroviridae Retroviridae - genetics RNA RNA, Messenger - metabolism RODENTS SOMATIC CELLS THERAPY TRANSCRIPTION Transcription, Genetic Transfection VERTEBRATES VIRUSES |
| title | Retroviral-Mediated Gene Transfer and Expression of Human Phenylalanine Hydroxylase in Primary Mouse Hepatocytes |
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