ArticleNucleic acids research2007
Comparison of RNAi efficiency mediated by tetracycline-responsive H1 and U6 promoter variants in mammalian cell lines.
Article in Nucleic acids research, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
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- A novel experimental approach for the selective isolation and characterization of human RNase MRP.RNA biology · 2022Article
- Review
- A simple approach for multi-targeted shRNA-mediated inducible knockdowns using Sleeping Beauty vectors.PloS one · 2018Article
- Optimized inducible shRNA and CRISPR/Cas9 platforms for in vitro studies of human development using hPSCs.Development (Cambridge, England) · 2016Article
- The Development of a Viral Mediated CRISPR/Cas9 System with Doxycycline Dependent gRNA Expression for Inducible In vitro and In vivo Genome Editing.Frontiers in molecular neuroscience · 2016Article
- Drug-inducible synergistic gene silencing with multiple small hairpin RNA molecules for gene function study in animal model.Transgenic research · 2015Article
- CRISPR transcriptional repression devices and layered circuits in mammalian cells.Nature methods · 2014Article
- Optimization and comparison of knockdown efficacy between polymerase II expressed shRNA and artificial miRNA targeting luciferase and Apolipoprotein B100.BMC biotechnology · 2012Article
- A versatile single-plasmid system for tissue-specific and inducible control of gene expression in transgenic mice.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2011Article
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- Conditional expression of retrovirally delivered anti-MYCN shRNA as an in vitro model system to study neuronal differentiation in MYCN-amplified neuroblastoma.BMC developmental biology · 2011Article
- Improved conditional expression systems resulting in physiological level of HNF4alpha expression confirm HNF4alpha induced apoptosis in the pancreatic beta-cell line INS-1.BMC research notes · 2009Article
- Efficient inhibition of human cytomegalovirus DNA polymerase expression by small hairpin RNA in vitro.Current microbiology · 2008Article
- AAV vectors for RNA-based modulation of gene expression.Gene therapy · 2008Review
- RNA interference and antiviral therapy.World journal of gastroenterology · 2007Review
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7 authors.
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Abstract
Conditional expression of short hairpin RNAs (shRNAs) to knock down target genes is a powerful tool to study gene function. The most common inducible expression systems are based on tetracycline-regulated RNA polymerase III promoters. During the last years, several tetracycline-inducible U6 and H1 promoter variants have been reported in different experimental settings showing variable efficiencies. In this study, we compare the most common variants of these promoters in several mammalian cell lines. For all cell lines tested, we find that several inducible U6 and H1 promoters containing single tetracycline operator (tetO) sequences show high-transcriptional background in the non-induced state. Promoter variants containing two tetO sequences show tight suppression of transcription in the non-induced state, and high tet responsiveness and high gene knockdown efficiency upon induction in all cell lines tested. We report a variant of the H1 promoter containing two O2-type tetO sequences flanking the TATA box that shows little transcriptional background in the non-induced state and up to 90% target knockdown when the inducer molecule (dox-doxycycline) is added. This inducible system for RNAi-based gene silencing is a good candidate for use both in basic research on gene function and for potential therapeutic applications.
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