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PMID: 7690960 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Production of high-titer helper-free retroviruses by transient transfection.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 18 ·1993-09-15 ·Pages 8392-6

Pear WS, Nolan GP, Scott ML, Baltimore D

Abstract

The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly transfectable 293T cell line into which are stably introduced constructs that express retroviral packaging functions. The resulting ecotropic virus packaging cell line BOSC 23 produces infectious retrovirus at > 10(6) infectious units/ml of supernatant within 72 hr after CaPO4-mediated transfection. A stringent assay for replication-competent virus showed that no helper virus was present. The system can produce high titers of retroviral vectors expressing genes that are extremely difficult to propagate at high titer in stable producer lines. This method should facilitate and extend the use of helper-free retroviral gene transfer, as well as be useful for gene therapy.

Related Genes

v-abl

MeSH Terms

3T3 Cells Animals Bone Marrow Transplantation/physiology Cell Line Cell Transformation, Neoplastic Chloroquine/pharmacology Female Genes, abl Helper Viruses/physiology Humans Male Mice Mice, Inbred C57BL Plasminogen Activators/biosynthesis RNA-Directed DNA Polymerase/analysis Repetitive Sequences, Nucleic Acid Retroviridae/drug effects,genetics,physiology Transfection Virus Replication/drug effects beta-Galactosidase/biosynthesis,metabolism

Chemicals

Chloroquine RNA-Directed DNA Polymerase beta-Galactosidase Plasminogen Activators

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Pear W S

Rockefeller University, New York, NY 10021.

Nolan G P

Scott M L

Baltimore D

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23 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.90.18.8392

Published

1993-09-15

Pages

8392-6

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC47362

Subset

Grants

NCI NIH HHS · CA51462 · United States

External Links

PubMed source DOI: 10.1073/pnas.90.18.8392 PMC Full Text View journal details

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