Toward a Comprehensive Temperature-Sensitive Mutant Repository of the Essential Genes of Saccharomyces cerevisiae

Shay Ben-Aroya; Candice Coombes; Teresa Kwok; Kathryn A. O'Donnell; Jef D. Boeke; Philip Hieter

doi:10.1016/j.molcel.2008.02.021

Toward a Comprehensive Temperature-Sensitive Mutant Repository of the Essential Genes of Saccharomyces cerevisiae

Shay Ben-Aroya, Candice Coombes, Teresa Kwok, Kathryn A. O'Donnell, Jef D. Boeke, Philip Hieter

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Abstract

The Saccharomyces cereivisiae gene deletion project revealed that approximately 20% of yeast genes are required for viability. The analysis of essential genes traditionally relies on conditional mutants, typically temperature-sensitive (ts) alleles. We developed a systematic approach (termed "diploid shuffle") useful for generating a ts allele for each essential gene in S. cerevisiae and for improved genetic manipulation of mutant alleles and gene constructs in general. Importantly, each ts allele resides at its normal genomic locus, flanked by specific cognate UPTAG and DNTAG bar codes. A subset of 250 ts mutants, including ts alleles for all uncharacterized essential genes and prioritized for genes with human counterparts, is now ready for distribution. The importance of this collection is demonstrated by biochemical and genetic screens that reveal essential genes involved in RNA processing and maintenance of chromosomal stability.

Original language	English
Pages (from-to)	248-258
Number of pages	11
Journal	Molecular Cell
Volume	30
Issue number	2
DOIs	https://doi.org/10.1016/j.molcel.2008.02.021
State	Published - 25 Apr 2008
Externally published	Yes

Bibliographical note

Funding Information:
We thank A. Carter, M. Tao, M. Kofoed, B. Le, and L. Tang for technical assistance, M. Kupiec, K. McManus, G. Simchen, and X. Pan for helpful discussions; and J. Woolford for helpful comments on an earlier version of the manuscript. Supported in part by National Institutes of Health (NIH) grant P01 CA16519 to P.H. and J.D.B., Canadian Institute for Health Research (CIHR) grant MOP-38096 to P.H., and NIH roadmap grant U54 RR020839 (to J.D.B.). S.B.-A. was supported by an Human Frontier Science Program (HFSP) long-term fellowship and by a research grant from the Killam trust.

Funding

We thank A. Carter, M. Tao, M. Kofoed, B. Le, and L. Tang for technical assistance, M. Kupiec, K. McManus, G. Simchen, and X. Pan for helpful discussions; and J. Woolford for helpful comments on an earlier version of the manuscript. Supported in part by National Institutes of Health (NIH) grant P01 CA16519 to P.H. and J.D.B., Canadian Institute for Health Research (CIHR) grant MOP-38096 to P.H., and NIH roadmap grant U54 RR020839 (to J.D.B.). S.B.-A. was supported by an Human Frontier Science Program (HFSP) long-term fellowship and by a research grant from the Killam trust.

Funders	Funder number
National Institutes of Health
National Cancer Institute	P01CA016519
National Center for Research Resources	U54RR020839
Canadian Institutes of Health Research	U54 RR020839, MOP-38096
Human Frontier Science Program
Killam Trusts

Keywords

DNA
RNA

Access to Document

10.1016/j.molcel.2008.02.021

Cite this

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title = "Toward a Comprehensive Temperature-Sensitive Mutant Repository of the Essential Genes of Saccharomyces cerevisiae",

abstract = "The Saccharomyces cereivisiae gene deletion project revealed that approximately 20% of yeast genes are required for viability. The analysis of essential genes traditionally relies on conditional mutants, typically temperature-sensitive (ts) alleles. We developed a systematic approach (termed {"}diploid shuffle{"}) useful for generating a ts allele for each essential gene in S. cerevisiae and for improved genetic manipulation of mutant alleles and gene constructs in general. Importantly, each ts allele resides at its normal genomic locus, flanked by specific cognate UPTAG and DNTAG bar codes. A subset of 250 ts mutants, including ts alleles for all uncharacterized essential genes and prioritized for genes with human counterparts, is now ready for distribution. The importance of this collection is demonstrated by biochemical and genetic screens that reveal essential genes involved in RNA processing and maintenance of chromosomal stability.",

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Toward a Comprehensive Temperature-Sensitive Mutant Repository of the Essential Genes of Saccharomyces cerevisiae

Abstract

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Keywords

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