Knocking down gene function with an RNA aptamer expressed as part of an intron.
We developed a powerful expression system to produce aptamers and other types of functional RNA in yeast to examine their effects. Utilizing the intron homing process, the aptamer-coding sequences were integrated into hundreds of rRNA genes, and the aptamers were transcribed at high levels by RNA polymerase I without any additional promoter being introduced into the cell. We used this system to express an aptamer against the heat shock factor 1 (HSF1), a conserved transcription factor responsible for mobilizing specific genomic expression programs in response to stressful conditions such as elevated temperature. We observed a temperature sensitive growth retardation phenotype and specific decrease of heat shock gene expression. As HSF1 enables and promotes malignant growth and metastasis in mammals, and this aptamer binds yeast HSF1 and its mammalian ortholog with equal affinity, the results presented here attest to the potential of this aptamer as a specific and effective inhibitor of HSF1 activity.
Wang
Shengchun
S
Department of Biological Sciences, University at Albany, State University of New York, Albany, NY 12222, USA.
Zhao
Xiaoching
X
Suran
Robert
R
Vogt
Volker M
VM
Lis
John T
JT
Shi
Hua
H
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D000076249:Heat Shock Transcription Factors; D007438:Introns; D010641:Phenotype; D012313:RNA / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014157:Transcription Factors / Q000037:antagonists & inhibitors*
D052157:Aptamers, Nucleotide; D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; D014157:Transcription Factors; D012313:RNA