Cited by 202Virus-based gene therapy researchParvovirus B19 Infection StudiesRNA Interference and Gene Delivery

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A Human Parvovirus, Adeno-Associated Virus, as a Eucaryotic Vector: Transient Expression and Encapsidation of the Procaryotic Gene for Chloramphenicol Acetyltransferase

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Tratschin, J D, West, M H, Sandbank, T, Carter, B J·1984·Molecular and Cellular Biology

DOI 10.1128/mcb.4.10.2072PMC PMC369024

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Abstract

We have used the defective human parvovirus adeno-associated virus (AAV) as a novel eucaryotic vector (parvector) for the expression of a foreign gene in human cells. The recombinant, pAV2, contains the AAV genome in a pBR322-derived bacterial plasmid. When pAV2 is transfected into human cells together with helper adenovirus particles, the AAV genome is rescued from the recombinant plasmid and replicated to produce infectious AAV particles at high efficiency. To create a vector, we inserted a procaryotic sequence coding for chloramphenicol acetyltransferase (CAT) into derivatives of pAV2 following either of the AAV promoters p40 (pAVHiCAT) and p19 (pAVBcCAT). When transfected into human 293 cells or HeLa cells, pAVHiCAT expressed CAT activity in the absence of adenovirus. In the presence of adenovirus, this vector produced increased amounts of CAT activity and the recombinant AAV-CAT genome was replicated. In 293 cells, pAVBcCAT expressed a similar amount of CAT activity in the absence or presence of adenovirus and the recombinant AAV-CAT genome was not replicated. In HeLa cells, pAVBcCAT expressed low levels of CAT activity, but this level was elevated by coinfection with adenovirus particles or by cotransfection with a plasmid which expressed the adenovirus early region 1A (E1A) product. The E1A product is a transcriptional activator and is expressed in 293 cells. Thus, expression from two AAV promoters is differentially regulated: expression from p19 is increased by E1A, w…

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Authors: Tratschin, J. D., West, M. H., Sandbank, T., & Carter, B. J.
Affiliations: National Institutes of Health (US)
Venue: Molecular and Cellular Biology
Publisher: Informa UK Limited
Published: 1984-10
Volume: 4
Issue: 10
Pages: 2072-2081
ISSN: 0270-7306 (ISSN-L)
1098-5549
DOI: 10.1128/mcb.4.10.2072
PMC: PMC369024
OpenAlex: W1824389368
Cited by: 202
References: 63
Citations by year: 2026: 2 · 2025: 5 · 2024: 7 · 2023: 5 · 2022: 3 · 2021: 6 · 2020: 14 · 2019: 4 · 2018: 4 · 2017: 6 · 2016: 3 · 2015: 2 · 2014: 6 · 2013: 3 · 2012: 3
FWCI: 3.75
Type: journal article
Language: en
Open access: green
europepmc.org
Locations: Molecular and Cellular Biology (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Health Sciences › Medicine › Infectious Diseases › Parvovirus B19 Infection Studies
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords: adeno-associated virus, parvovirus, chloramphenicol acetyltransferase, transient expression, viral encapsidation, recombinant adeno-associated virus, 293 cells, HeLa cells
MeSH terms: Acetyltransferases, Chimera, DNA, Viral, Gene Expression Regulation, Genetic Vectors, HeLa Cells, Parvoviridae, Plasmids, Recombination, Genetic, Transfection, Chloramphenicol O-Acetyltransferase

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A Human Parvovirus, Adeno-Associated Virus, as a Eucaryotic Vector: Transient Expression and Encapsidation of the Procaryotic Gene for Chloramphenicol Acetyltransferase

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