High efficiency transformation by direct microinjection of DNA into cultured mammalian cells.

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Publication Type Journal Article
School or College College of Science; School of Medicine
Department Oncological Sciences; Biology; Human Genetics
Program Institute of Human Genetics; Howard Hughes Medical Institute (HHMI)
Creator Capecchi, Mario R.
Title High efficiency transformation by direct microinjection of DNA into cultured mammalian cells.
Date 1980-11-22
Description Direct microinjection of DNA by glass micropipettes was used to introduce the Herpes simplex virus thymidine kinase gene into cultured mammalian cells. When DNA was delivered directly into the nuclei of LMTK-, a mouse cell line deficient in thymidine kinase activity, 50--100% of the cells expressed TK enzymatic activity. In contrast, no TK activity could be detected when the DNA was injected into the cytoplasm. The number of injected LMTK- cells capable of indefinite growth in a TK+ selective medium (that is, transformants) depended on the nature of the plasmid DNA into which the HSV-TK gene was inserted. One cell in 500-1000 cells which received nuclear injections with pBR322/TK DNA gave rise to a viable colony when grown in HAT medium (that is, a TK+ selective medium). The transformation frequency increased to one in five injected cells when specific SV40 DNA sequences were also introduced into the HSV-TK plasmid. With the microinjection procedure transformation frequency was relatively insensitive to DNA concentration and did not depend on co-injecting with a carrier DNA. Most of the transformants were stable in nonselective medium as soon as they could be tested.
Type Text
Publisher MIT
Volume 22
Issue 2 Pt 2
First Page 479
Last Page 488
Subject Cell Nucleus; Cytoplasm; DNA, Viral; Microinjections; Recombination, Genetic
Subject MESH Cells, Cultured; Thymidine Kinase; Transformation, Genetic
Language eng
Bibliographic Citation Cell. 1980 Nov;22(2 Pt 2):479-88: Capecchi MR. High efficiency transformation by direct microinjection of DNA into cultured mammalian cells. Retrieved on September 25.2006 from
Rights Management Copyright © 1980, MIT. All Rights Reserved.
Format Medium application/pdf
Identifier ir-main,469
ARK ark:/87278/s63f572m
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2021-05-06
ID 704814
Reference URL https://collections.lib.utah.edu/ark:/87278/s63f572m
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