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PMID: 6952193 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mechanisms of DNA uptake by mammalian cells: fate of exogenously added DNA monitored by the use of fluorescent dyes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 2 ·1982-01-00 ·Pages 422-6

Loyter A, Scangos GA, Ruddle FH

Abstract

Coprecipitation of DNA with calcium phosphate is a commonly used method of gene transfer in mammalian cells. We have found that DNA forms a tight complex with Ca Pi and that DNA in this complex is resistant to nucleases present in serum or added externally. Under optimal conditions, virtually all of the recipient mouse Ltk- Aprt- cells take up Ca Pi--DNA complexes, as determined by fluorescent dyes specific for DNA (4',6-diaminilo-2-phenylindol dihydrochloride) or for calcium salts (chlorotetracycline). However, only a small proportion of the cells have detectable CA Pi--DNA complexes in the nucleus. Uptake of the Ca Pi--DNA complexes was highly dependent upon the pH at which the Ca Pi--DNA complexes was formed and upon the concentration of DNA in the complex.

MeSH Terms

Animals Biological Transport Calcium Phosphates Cell Line Chemical Precipitation Chlortetracycline DNA/metabolism Fluorescent Dyes Hydrogen-Ion Concentration Indoles Mice Transformation, Genetic

Chemicals

Calcium Phosphates Fluorescent Dyes Indoles DAPI DNA Chlortetracycline

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Loyter A

Scangos G A

Ruddle F H

References (14)

14 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.79.2.422

Published

1982-01-00

Pages

422-6

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC345755

Subset

Grants

NIGMS NIH HHS · GM09966 · United States

External Links

PubMed source DOI: 10.1073/pnas.79.2.422 PMC Full Text View journal details

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Mechanisms of DNA uptake by mammalian cells: fate of exogenously added DNA monitored by the use of fluorescent dyes.

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