Design and Structural Analysis of Hairpin-TFO for Transcriptional Activation of Genes in S. cerevisiae

Triplex forming oligonucleotides (TFOs) have the potential to modulate gene expression. While most of the experiments are directed towards triplex mediated inhibition of gene expression the strategy potentially could be used for gene specific activation. In an attempt to design a strategy for gene s...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biomolecular structure & dynamics 2002-10, Vol.20 (2), p.265-273
Main Authors: Ghosh, Mrinal Kanti, Katyal, Anju, Brahmachari, Vani, Chandra, Ramesh
Format: Article
Language:English
Subjects:
Base Sequence
Binding Sites
Chromosomes, Fungal - genetics
Circular Dichroism
DNA, Fungal - metabolism
DNA-Binding Proteins - metabolism
Electrophoretic Mobility Shift Assay (EMSA)
Gal4
Gene Expression Regulation - drug effects
Genes, Fungal
Hairpin-TFO
Models, Biological
Nucleic Acid Heteroduplexes
Oligonucleotides - chemical synthesis
Oligonucleotides - metabolism
Polyethyleneimine - pharmacology
Polypurine/Polypyrimidine
Promoter Regions, Genetic
Protein Binding
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae Proteins - metabolism
Temperature
Transcription Activation
Transcription Factors - metabolism
Transcription, Genetic
Transcriptional Activation
Triple helix
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Triplex forming oligonucleotides (TFOs) have the potential to modulate gene expression. While most of the experiments are directed towards triplex mediated inhibition of gene expression the strategy potentially could be used for gene specific activation. In an attempt to design a strategy for gene specific activation in vivo applicable to a large number of genes we have designed a TFO based activator-target system which may be utilized in Saccharomyces cerevisiae or any other system where Ga14 protein is ectopically expressed. The total genome sequence of Saccharomyces cerevisiae and expression profiles were used to select the target genes with upstream poly (pu/py) sequences. We have utilized the paradigm of Gal4 protein and its binding site. We describe here the selection of target genes and design of hairpin-TFO including the targeting sequences containing polypurine stretch found in the upstream promoter regions of weakly expressed genes. We demonstrate, the formation of hairpin-TFO, its binding to Gal4 protein, its ability to form triplex with the target duplex in vitro, the effect of polyethylenimine on complex formation and discuss the implication on in vivo transcription activation.
ISSN:0739-1102
1538-0254
DOI:10.1080/07391102.2002.10506842