Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology

Xudong Cheng, Jae Hyeong Ko, Sidney Altman

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The artificial inhibition of expression of genes in Saccharomyces cerevisiae is not a widespread, useful phenomenon. The external guide sequence (EGS) technology, which is well-proven in bacteria and mammalian cells in tissue culture and in mice, can also be utilized in yeast. The TOP2 and SRG1 genes can be inhibited by ∼30% with EGSs in vivo. Results in vitro also show convenient cleavage of the relevant transcripts by RNase P and appropriate EGSs. The feasible constructs shown to date have an EGS covalently linked to M1 RNA, the RNA subunit of RNase P from Escherichia coli. Greater efficiency in cleavage of transcripts can be fashioned using more than one EGS targeted to different sites in a transcript and stronger promoters controlling the EGS constructs. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)544-549
Number of pages6
JournalRNA
Volume17
Issue number3
DOIs
StatePublished - Mar 1 2011
Externally publishedYes

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Keywords

  • RNase P
  • SRG1 gene
  • TOP2 gene
  • Target RNA

ASJC Scopus subject areas

  • Molecular Biology

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