- THIS ARTICLE
- Full Text
- Full Text (PDF)
-
All Versions of this Article:
genetics.106.056945v1
173/2/769 most recent - Alert me when this article is cited
- Alert me if a correction is posted
- SERVICES
- Email this article to a friend
- Similar articles in this journal
- Similar articles in PubMed
- Alert me to new issues of the journal
- Download to citation manager
- Reprints & Permissions
- CITING ARTICLES
- Citing Articles via HighWire
- Citing Articles via Google Scholar
- GOOGLE SCHOLAR
- Articles by Bateman, J. R.
- Articles by Wu, C.-t.
- Search for Related Content
- PUBMED
- PubMed Citation
- Articles by Bateman, J. R.
- Articles by Wu, C.-t.
Originally published as Genetics Published Articles Ahead of Print on March 17, 2006.
Genetics, Vol. 173, 769-777, June 2006, Copyright © 2006
doi:10.1534/genetics.106.056945
Site-Specific Transformation of Drosophila via 
C31 Integrase-Mediated Cassette Exchange
Jack R. Bateman*,1,
Anne M. Lee*,1 and
C.-ting Wu*,
,2
* Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115 and Division of Genetics and Division of Molecular Medicine, Harvard Medical School, Boston, Massachusetts 02115
2 Corresponding author: Harvard Medical School, Department of Genetics, 77 Avenue Louis Pasteur, New Research Building, Room 264, Boston, MA 02115.
E-mail: twu{at}genetics.med.harvard.edu
Position effects can complicate transgene analyses. This is especially true when comparing transgenes that have inserted randomly into different genomic positions and are therefore subject to varying position effects. Here, we introduce a method for the precise targeting of transgenic constructs to predetermined genomic sites in Drosophila using the 
C31 integrase system in conjunction with recombinase-mediated cassette exchange (RMCE). We demonstrate the feasibility of this system using two donor cassettes, one carrying the yellow gene and the other carrying GFP. At all four genomic sites tested, we observed exchange of donor cassettes with an integrated target cassette carrying the mini-white gene. Furthermore, because RMCE-mediated integration of the donor cassette is necessarily accompanied by loss of the target cassette, we were able to identify integrants simply by the loss of mini-white eye color. Importantly, this feature of the technology will permit integration of unmarked constructs into Drosophila, even those lacking functional genes. Thus, C31 integrase-mediated RMCE should greatly facilitate transgene analysis as well as permit new experimental designs.
This article has been cited by other articles:
 |
|
 |
|
  G. Gao, N. Wesolowska, and Y. S. Rong SIRT Combines Homologous Recombination, Site-Specific Integration, and Bacterial Recombineering for Targeted Mutagenesis in Drosophila CSH Protocols, June 1, 2009; 2009(6): pdb.prot5236 - pdb.prot5236. [Abstract] [Full Text]
|
|
|
|
 |
|
 J. Huang, W. Zhou, W. Dong, A. M. Watson, and Y. Hong From the Cover: Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering PNAS, May 19, 2009; 106(20): 8284 - 8289. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. M. Choi, S. Vilain, M. Langen, S. Van Kelst, N. De Geest, J. Yan, P. Verstreken, and B. A. Hassan Conditional Mutagenesis in Drosophila Science, April 3, 2009; 324(5923): 54 - 54. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Jia, Y. Liu, W. Yan, and J. Jia PP4 and PP2A regulate Hedgehog signaling by controlling Smo and Ci phosphorylation Development, January 15, 2009; 136(2): 307 - 316. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Bateman and C.-t. Wu A Simple Polymerase Chain Reaction-Based Method for the Construction of Recombinase-Mediated Cassette Exchange Donor Vectors Genetics, November 1, 2008; 180(3): 1763 - 1766. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Thyagarajan, Y. Liu, S. Shin, U. Lakshmipathy, K. Scheyhing, H. Xue, C. Ellerstrom, R. Strehl, J. Hyllner, M. S. Rao, et al. Creation of Engineered Human Embryonic Stem Cell Lines Using phiC31 Integrase Stem Cells, January 1, 2008; 26(1): 119 - 126. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. J. T. Venken and H. J. Bellen Transgenesis upgrades for Drosophila melanogaster Development, October 15, 2007; 134(20): 3571 - 3584. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Gupta, R. Till, and M. C. M. Smith Sequences in attB that affect the ability of {phi}C31 integrase to synapse and to activate DNA cleavage Nucleic Acids Res., May 11, 2007; 35(10): 3407 - 3419. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Bischof, R. K. Maeda, M. Hediger, F. Karch, and K. Basler An optimized transgenesis system for Drosophila using germ-line-specific {varphi}C31 integrases PNAS, February 27, 2007; 104(9): 3312 - 3317. [Abstract] [Full Text] [PDF]
|
|