NDLI: Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells.

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Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Jasin, M. Rouet, P. Smih, F.
Description	Author Affiliation: Rouet P ( Cell Biology and Genetics Program, Sloan-Kettering Institute, New York, NY 10021.);
Abstract	Double-strand breaks introduced into DNA in vivo have been shown to enhance homologous recombination in a variety of chromosomal and extrachromosomal loci in Saccharomyces cerevisiae. To introduce double-strand breaks in DNA at defined locations in mammalian cells, we have constructed a mammalian expression vector for a modified form of I-Sce I, a yeast mitochondrial intron-encoded endonuclease with an 18-bp recognition sequence. Expression of the modified I-Sce I endonuclease in COS1 cells results in cleavage of model recombination substrates and enhanced extrachromosomal recombination, as assayed by chloramphenicol acetyltransferase activity and Southern blot analysis. Constitutive expression of the endonuclease in mouse 3T3 cells is not lethal, possibly due to either the lack of I-Sce I sites in the genome or sufficient repair of them. Expression of an endonuclease with such a long recognition sequence will provide a powerful approach to studying a number of molecular processes in mammalian cells, including homologous recombination.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	13
Volume Number	91
Language	English
Publisher	National Academy of Sciences
Publisher Date	1994-07-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA Restriction Enzymes Metabolism Recombination, Genetic 3T3 Cells Animals Avian Sarcoma Viruses Genetics Binding Sites Blotting, Southern Cell Line Cercopithecus Aethiops Chloramphenicol O-Acetyltransferase Biosynthesis DNA Chemistry Deoxyribonucleases, Type II Site-Specific Escherichia Coli Kidney Mice Molecular Sequence Data Oligodeoxyribonucleotides Plasmids Promoter Regions, Genetic Restriction Mapping Transfection Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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