Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology
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 gene...
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| Veröffentlicht in: | RNA (Cambridge) 2011-03, Vol.17 (3), p.544-549 |
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| creator | Cheng, Xudong Ko, Jae-Hyeong Altman, Sidney |
| description | 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. |
| doi_str_mv | 10.1261/rna.2538711 |
| format | Article |
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| subjects | Blotting, Northern Escherichia coli Escherichia coli - enzymology Escherichia coli - genetics Method Ribonuclease P - metabolism RNA, Fungal - genetics RNA, Fungal - metabolism RNA, Guide - chemistry RNA, Guide - pharmacology RNA, Messenger - genetics RNA, Messenger - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - growth & development Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins - antagonists & inhibitors Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism |
| title | Inactivation of expression of two genes in Saccharomyces cerevisiae with the external guide sequence methodology |
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