Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (148K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Simons, A. H.
Right arrow Articles by Canaani, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Simons, A. H.
Right arrow Articles by Canaani, D.
Related Collections
Right arrow Cell biology
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 20 e100
© 2001 Oxford University Press

Genetic synthetic lethality screen at the single gene level in cultured human cells

Arnold H. Simons, Naomi Dafni, Iris Dotan, Yoram Oron1 and Dan Canaani*

Department of Biochemistry,Sherman Building, Room 604, The George S. Wise Faculty of Life Sciences and 1Department of Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, Ramat Aviv, 69978 Israel

Recently, we demonstrated the feasibility of a chemical synthetic lethality screen in cultured human cells. We now demonstrate the principles for a genetic synthetic lethality screen. The technology employs both an immortalized human cell line deficient in the gene of interest, which is complemented by an episomal survival plasmid expressing the wild-type cDNA for the gene of interest, and the use of a novel GFP-based double-label fluorescence system. Dominant negative genetic suppressor elements (GSEs) are selected from an episomal library expressing short truncated sense and antisense cDNAs for a gene likely to be synthetic lethal with the gene of interest. Expression of these GSEs prevents spontaneous loss of the GFP-marked episomal survival plasmid, thus allowing FACS enrichment for cells retaining the survival plasmid (and the GSEs). The dominant negative nature of the GSEs was validated by the decreased resident enzymatic activity present in cells harboring the GSEs. Also, cells mutated in the gene of interest exhibit reduced survival upon GSE expression. The identification of synthetic lethal genes described here can shed light on functional genetic interactions between genes involved in normal cell metabolism and in disease.

* To whom correspondence should be addressed. Tel: +972 3 6408985; Fax: +972 3 6424270; Email: canaani{at}post.tau.ac.il


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Molecular Cancer TherapeuticsHome page
Y. Einav, E. Shistik, M. Shenfeld, A. H. Simons, D. W. Melton, and D. Canaani
Replication and episomal maintenance of Epstein-Barr virus-based vectors in mouse embryonal fibroblasts enable synthetic lethality screens
Mol. Cancer Ther., November 1, 2003; 2(11): 1121 - 1128.
[Abstract] [Full Text] [PDF]

Home page
 J Biomol ScreenHome page
M. Pierce, C. Wang, M. Rebentisch, M. Endo, M. Stump, and A. Kamb
A High-Throughput, Homogeneous Microplate Assay for Agents That Kill Mammalian Tissue Culture Cells
J Biomol Screen, June 1, 2003; 8(3): 283 - 291.
[Abstract] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.