Nucleic Acids Research, 1995, Vol. 23, No. 24 5012-5019
© 1995
Articles |
Double-strand breaks at the target locus stimulate gene targeting in embryonic stem cells
Cell Biology and Genetics Program New York, NY 10021, USA 1Molecular Biology Program, Sloan-Kettering Institute and Comell University Graduate School of Medical Sciences New York, NY 10021, USA
*To whom correspondence should be addressed
Received September 14, 1995. Accepted November 6, 1995.
Double-strand breaks (DSBs) are recomblnogenlc lesions In chromosomal DNA in yeast, Drosophlla and Caenortiabdltls elegans. Recent studies In mammalian cells utilizing the l-Scel endonuclease have demonstrated that in some immortalized cell lines DSBs In chromosomal DNA are also recomblnogenic. We have now tested embryonic stem (ES) cells, a non-transformed mouse cell line frequently used in gene targeting studies. We find that a DSB introduced by l-Scel stimulates gene targeting at a selectable neo locus at least 50-fold. The enhanced level of targeting is achieved by transient expression of the I-Sce) endonuclease. In 97% of targeted clones a single base pair polymorphism in the transfected homologous fragment was incorporated into the target locus. Analysis of the targeted locus demonstrated that most of the homologous recombination events were ‘twosided’, in contrast to previous studies In 3T3 cells in which ‘one-sided’ homologous events predominated. Thus ES cells may be more faithful in incorporating homologous fragments into their genome than other cells in culture.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  Z. Chen, F. Wen, N. Sun, and H. Zhao Directed evolution of homing endonuclease I-SceI with altered sequence specificity Protein Eng. Des. Sel., April 1, 2009; 22(4): 249 - 256. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Beumer, G. Bhattacharyya, M. Bibikova, J. K. Trautman, and D. Carroll Efficient Gene Targeting in Drosophila With Zinc-Finger Nucleases Genetics, April 1, 2006; 172(4): 2391 - 2403. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Chames, J.-C. Epinat, S. Guillier, A. Patin, E. Lacroix, and F. Paques In vivo selection of engineered homing endonucleases using double-strand break induced homologous recombination Nucleic Acids Res., November 23, 2005; 33(20): e178 - e178. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. L. Posey, V. Koufopanou, A. Burt, and F. S. Gimble Evolution of divergent DNA recognition specificities in VDE homing endonucleases from two yeast species Nucleic Acids Res., July 27, 2004; 32(13): 3947 - 3956. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Egli, E. Hafen, and W. Schaffner An Efficient Method to Generate Chromosomal Rearrangements by Targeted DNA Double-Strand Breaks in Drosophila melanogaster Genome Res., July 1, 2004; 14(7): 1382 - 1393. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. G. Miller, L. M. Petek, and D. W. Russell Human Gene Targeting by Adeno-Associated Virus Vectors Is Enhanced by DNA Double-Strand Breaks Mol. Cell. Biol., May 15, 2003; 23(10): 3550 - 3557. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. H. Porteus, T. Cathomen, M. D. Weitzman, and D. Baltimore Efficient Gene Targeting Mediated by Adeno-Associated Virus and DNA Double-Strand Breaks Mol. Cell. Biol., May 15, 2003; 23(10): 3558 - 3565. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Bibikova, K. Beumer, J. K. Trautman, and D. Carroll Enhancing Gene Targeting with Designed Zinc Finger Nucleases Science, May 2, 2003; 300(5620): 764 - 764. [Full Text] [PDF]
|
|
|
|
 |
|
 K. M. Vasquez, K. Marburger, Z. Intody, and J. H. Wilson Manipulating the mammalian genome by homologous recombination PNAS, July 17, 2001; 98(15): 8403 - 8410. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Elliott and M. Jasin Repair of Double-Strand Breaks by Homologous Recombination in Mismatch Repair-Defective Mammalian Cells Mol. Cell. Biol., April 15, 2001; 21(8): 2671 - 2682. [Abstract] [Full Text]
|
|
|
|
|
|
C. Richardson and M. Jasin Coupled Homologous and Nonhomologous Repair of a Double-Strand Break Preserves Genomic Integrity in Mammalian Cells Mol. Cell. Biol., December 1, 2000; 20(23): 9068 - 9075. [Abstract] [Full Text]
|
|
|
|
|
|
Z. Luo, M. A. Macris, A. F. Faruqi, and P. M. Glazer High-frequency intrachromosomal gene conversion induced by triplex-forming oligonucleotides microinjected into mouse cells PNAS, July 12, 2000; (2000) 160004997. [Abstract] [Full Text]
|
|
|
|
|
|
C. N. Sprung, G. E. Reynolds, M. Jasin, and J. P. Murnane Chromosome healing in mouse embryonic stem cells PNAS, June 8, 1999; 96(12): 6781 - 6786. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Richardson, M. E. Moynahan, and M. Jasin Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations Genes & Dev., December 15, 1998; 12(24): 3831 - 3842. [Abstract] [Full Text]
|
|
|
|
|
|
G. Donoho, M. Jasin, and P. Berg Analysis of Gene Targeting and Intrachromosomal Homologous Recombination Stimulated by Genomic Double-Strand Breaks in Mouse Embryonic Stem Cells Mol. Cell. Biol., July 1, 1998; 18(7): 4070 - 4078. [Abstract] [Full Text]
|
|
|
|
|
|
F. Liang, M. Han, P. J. Romanienko, and M. Jasin Homology-directed repair is a major double-strand break repair pathway in mammalian cells PNAS, April 28, 1998; 95(9): 5172 - 5177. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Cohen-Tannoudji, S. Robine, A. Choulika, D. Pinto, F. El Marjou, C. Babinet, D. Louvard, and F. Jaisser I-SceI-Induced Gene Replacement at a Natural Locus in Embryonic Stem Cells Mol. Cell. Biol., March 1, 1998; 18(3): 1444 - 1448. [Abstract] [Full Text]
|
|
|
|
|
|
B. Elliott, C. Richardson, J. Winderbaum, J. A. Nickoloff, and M. Jasin Gene Conversion Tracts from Double-Strand Break Repair in Mammalian Cells Mol. Cell. Biol., January 1, 1998; 18(1): 93 - 101. [Abstract] [Full Text]
|
|
|
|
|
|
M. E. Moynahan and M. Jasin Loss of heterozygosity induced by a chromosomal double-strand break PNAS, August 19, 1997; 94(17): 8988 - 8993. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Luo, M. A. Macris, A. F. Faruqi, and P. M. Glazer High-frequency intrachromosomal gene conversion induced by triplex-forming oligonucleotides microinjected into mouse cells PNAS, August 1, 2000; 97(16): 9003 - 9008. [Abstract] [Full Text] [PDF]
|
|