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Nucleic Acids Research, 2000, Vol. 28, No. 6 E15-e15
© 2000 Oxford University Press

A rapid genetic screening system for identifying gene-specific suppression constructs for use in human cells

Greg M. Arndt*, Margaret Patrikakis and David Atkins

Johnson & Johnson Research, The Australian Technology Park, Level 4, 1 Central Avenue, Eveleigh, NSW 1430, Sydney, Australia

We describe a rapid cell-based genetic screen using fission yeast for identifying efficient gene suppression constructs (GSCs) from large libraries (105) for any target sequence for use in human cells. In this system, target sequences are fused to the 5' end of the lacZ reporter gene and expressed in yeast. Random fragment expression libraries derived from the target sequence are screened in the fusion gene-expressing strain using the lacZ gene-encoded colony color phenotype. We demonstrate the utility of this screening assay by identifying a range of different GSCs for the fission yeast ura4 gene and human c-myc and Chk1 sequences, including rare efficient suppressors. GSCs specific for c-myc were shown to regulate expression of both a c-myc–lacZ fusion gene and the endogenous c-myc gene in human cells.

* To whom correspondence should be addressed at: Johnson & Johnson Research, The Australian Technology Park, Locked Bag 4555, Strawberry Hills, NSW 2012, Sydney, Australia. Tel: +61 2 8396 5837; Fax: +61 2 8396 5811; Email: garndt@medau.jnj.com


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