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 Citations (Scopus)

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 2011
Externally publishedYes

Fingerprint

Ribonuclease P
Saccharomyces cerevisiae
RNA
Gene Expression
Yeasts
Escherichia coli
Technology
Bacteria
Genes
In Vitro Techniques

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology. / Cheng, Xudong; Ko, Jae Hyeong; Altman, Sidney.

In: RNA, Vol. 17, No. 3, 03.2011, p. 544-549.

Research output: Contribution to journalArticle

@article{c1772029ffda42d788fba3b2285ab64c,
title = "Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology",
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.",
keywords = "RNase P, SRG1 gene, Target RNA, TOP2 gene",
author = "Xudong Cheng and Ko, {Jae Hyeong} and Sidney Altman",
year = "2011",
month = "3",
doi = "10.1261/rna.2538711",
language = "English (US)",
volume = "17",
pages = "544--549",
journal = "RNA",
issn = "1355-8382",
publisher = "Cold Spring Harbor Laboratory Press",
number = "3",

}

TY - JOUR

T1 - Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology

AU - Cheng, Xudong

AU - Ko, Jae Hyeong

AU - Altman, Sidney

PY - 2011/3

Y1 - 2011/3

N2 - 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.

AB - 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.

KW - RNase P

KW - SRG1 gene

KW - Target RNA

KW - TOP2 gene

UR - http://www.scopus.com/inward/record.url?scp=79951541189&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79951541189&partnerID=8YFLogxK

U2 - 10.1261/rna.2538711

DO - 10.1261/rna.2538711

M3 - Article

VL - 17

SP - 544

EP - 549

JO - RNA

JF - RNA

SN - 1355-8382

IS - 3

ER -