This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hasty, P Articles by Bradley, A Search for Related Content PubMed PubMed Citation Articles by Hasty, P Articles by Bradley, A

 Previous Article  |  Next Article 

Mol Cell Biol. 1991 November; 11(11): 5586-5591

The length of homology required for gene targeting in embryonic stem cells.

Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.

ABSTRACT

Homologous recombination has been used to introduce site-specific mutations into murine embryonic stem (ES) cells with both insertion and replacement vectors. In this study, we compared the frequency of gene targeting with various lengths of homology and found a dramatic increase in targeting with an increase in homology from 1.3 to 6.8 kb. We examined in detail the relationship between the length of homology and the gene-targeting frequency for replacement vectors and found that a critical length of homology is needed for targeting. Adding greater lengths of homology to this critical length has less of an effect on the targeting frequency. We also analyzed the lengths of homology necessary on both arms of the vector for gene replacement events and found that 472 bp of homology is used as efficiently as 1.2 kb in the formation and resolution of crossover junctions.

This article has been cited by other articles:

• van Zyl, L. J., van Munster, J. M., Rawlings, D. E. (2008). Construction of arsB and tetH Mutants of the Sulfur-Oxidizing Bacterium Acidithiobacillus caldus by Marker Exchange. Appl. Environ. Microbiol. 74: 5686-5694 [Abstract] [Full Text]  
• Adams, D. J., van der Weyden, L. (2008). Contemporary approaches for modifying the mouse genome. Physiol. Genomics 34: 225-238 [Abstract] [Full Text]  
• Barnes, R. L., McCulloch, R. (2007). Trypanosoma brucei homologous recombination is dependent on substrate length and homology, though displays a differential dependence on mismatch repair as substrate length decreases. Nucleic Acids Res 35: 3478-3493 [Abstract] [Full Text]  
• Boissinot, S., Davis, J., Entezam, A., Petrov, D., Furano, A. V. (2006). Fitness cost of LINE-1 (L1) activity in humans. Proc. Natl. Acad. Sci. USA 103: 9590-9594 [Abstract] [Full Text]  
• Nakayama, K., Ishida, T. (2006). Alu-mediated 100-kb deletion in the primate genome: The loss of the agouti signaling protein gene in the lesser apes.. Genome Res 16: 485-490 [Abstract] [Full Text]  
• McCulloch, R. D., Baker, M. D. (2006). Analysis of one-sided marker segregation patterns resulting from Mammalian gene targeting.. Genetics 172: 1767-1781 [Abstract] [Full Text]  
• Kamisugi, Y., Cuming, A. C., Cove, D. J. (2005). Parameters determining the efficiency of gene targeting in the moss Physcomitrella patens. Nucleic Acids Res 33: e173-e173 [Abstract] [Full Text]  
• Aoyama, M., Agari, K., Sun-Wada, G.-H., Futai, M., Wada, Y. (2005). Simple and straightforward construction of a mouse gene targeting vector using in vitro transposition reactions. Nucleic Acids Res 33: e52-e52 [Abstract] [Full Text]  
• Chen, Y.-T., Levasseur, R., Vaishnav, S., Karsenty, G., Bradley, A. (2004). Bigenic Cre/loxP, pu{Delta}tk conditional genetic ablation. Nucleic Acids Res 32: e161-e161 [Abstract] [Full Text]  
• Chen, Y.-T., Liu, P., Bradley, A. (2004). Inducible Gene Trapping with Drug-Selectable Markers and Cre/loxP To Identify Developmentally Regulated Genes. Mol. Cell. Biol. 24: 9930-9941 [Abstract] [Full Text]  
• Birmingham, E. C., Lee, S. A., McCulloch, R. D., Baker, M. D. (2004). Testing Predictions of the Double-Strand Break Repair Model Relating to Crossing Over in Mammalian Cells. Genetics 168: 1539-1555 [Abstract] [Full Text]  
• Lu, Z. H., Books, J. T., Kaufman, R. M., Ley, T. J. (2003). Long targeting arms do not increase the efficiency of homologous recombination in the {beta}-globin locus of murine embryonic stem cells. Blood 102: 1531-1533 [Abstract] [Full Text]  
• Seum, C., Pauli, D., Delattre, M., Jaquet, Y., Spierer, A., Spierer, P. (2002). Isolation of Su(var)3-7 Mutations by Homologous Recombination in Drosophila melanogaster. Genetics 161: 1125-1136 [Abstract] [Full Text]  
• Vasquez, K. M., Marburger, K., Intody, Z., Wilson, J. H. (2001). Manipulating the mammalian genome by homologous recombination. Proc. Natl. Acad. Sci. USA 98: 8403-8410 [Abstract] [Full Text]  
• Boissinot, S., Entezam, A., Furano, A. V. (2001). Selection Against Deleterious LINE-1-Containing Loci in the Human Lineage. Mol Biol Evol 18: 926-935 [Abstract] [Full Text]  
• Dragatsis, I., Zeitlin, S. (2001). A method for the generation of conditional gene repair mutations in mice. Nucleic Acids Res 29: e10-e10 [Abstract] [Full Text]  
• Li, J., Baker, M. D. (2000). Mechanisms Involved in Targeted Gene Replacement in Mammalian Cells. Genetics 156: 809-821 [Abstract] [Full Text]  
• Yesland, K., Fonzi, W. A. (2000). Allele-specific gene targeting in Candida albicans results from heterology between alleles. Microbiology 146: 2097-2104 [Abstract] [Full Text]  
• Akiyama, K., Watanabe, H., Tsukada, S., Sasai, H. (2000). A novel method for constructing gene-targeting vectors. Nucleic Acids Res 28: e77-e77 [Abstract] [Full Text]  
• Casqueiro, J., Gutiérrez, S., Bañuelos, O., Hijarrubia, M. J., Martín, J. F. (1999). Gene Targeting in Penicillium chrysogenum: Disruption of the lys2 Gene Leads to Penicillin Overproduction. J. Bacteriol. 181: 1181-1188 [Abstract] [Full Text]  
• Cohen-Tannoudji, M., Robine, S., Choulika, A., Pinto, D., El Marjou, F., Babinet, C., Louvard, D., Jaisser, F. (1998). I-SceI-Induced Gene Replacement at a Natural Locus in Embryonic Stem Cells. Mol. Cell. Biol. 18: 1444-1448 [Abstract] [Full Text]  
• Watson, J E, Slorach, E M, Maule, J, Lawson, D, Porteous, D J, Brookes, A J (1995). Human repeat-mediated integration of selectable markers into somatic cell hybrids.. Genome Res 5: 444-452 [Abstract]