ArticleProceedings of the National Academy of Sciences of the United States of America2005
RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 48 papers.
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Who cites it
48 citing papers in PubMed, 91 citations in OpenAlex.
- ESI mutagenesis: a one-step method for introducing mutations into bacterial artificial chromosomes.Life science alliance · 2021Article
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- Multimerization properties of PiggyMac, a domesticated piggyBac transposase involved in programmed genome rearrangements.Nucleic acids research · 2017Article
- Type I γ Phosphatidylinositol Phosphate 5-Kinase i5 Controls the Ubiquitination and Degradation of the Tumor Suppressor Mitogen-inducible Gene 6.The Journal of biological chemistry · 2016Article
- HP1BP3, a Chromatin Retention Factor for Co-transcriptional MicroRNA Processing.Molecular cell · 2016Article
- Minireview: Familiar Faces in Unfamiliar Places: The Emerging Role of Nuclear Receptors in Lung Cancer.Molecular endocrinology (Baltimore, Md.) · 2015Review
- RAS Synthetic Lethal Screens Revisited: Still Seeking the Elusive Prize?Clinical cancer research : an official journal of the American Association for Cancer Research · 2015Review
- Affinity proteomics to study endogenous protein complexes: pointers, pitfalls, preferences and perspectives.BioTechniques · 2015Article
- SNW1 enables sister chromatid cohesion by mediating the splicing of sororin and APC2 pre-mRNAs.The EMBO journal · 2014Article
- RNAi screening comes of age: improved techniques and complementary approaches.Nature reviews. Molecular cell biology · 2014Review
- Response to Shuda et al.The Journal of investigative dermatology · 2014Article
- A genomic toolkit to investigate kinesin and myosin motor function in cells.Nature cell biology · 2013Article
- Subdiffraction imaging of centrosomes reveals higher-order organizational features of pericentriolar material.Nature cell biology · 2012Article
- CEP192 interacts physically and functionally with the K63-deubiquitinase CYLD to promote mitotic spindle assembly.Cell cycle (Georgetown, Tex.) · 2012Article
- Molecular biology. Use and abuse of RNAi to study mammalian gene function.Science (New York, N.Y.) · 2012Article
- Improved methodology for the affinity isolation of human protein complexes expressed at near endogenous levels.BioTechniques · 2012Article
- Dynamic reorganization of Eg5 in the mammalian spindle throughout mitosis requires dynein and TPX2.Molecular biology of the cell · 2012Article
- From RNAi screens to molecular function in embryonic stem cells.Stem cell reviews and reports · 2012Review
- Triple SILAC to determine stimulus specific interactions in the Wnt pathway.Journal of proteome research · 2012Article
- High-efficiency counterselection recombineering for site-directed mutagenesis in bacterial artificial chromosomes.Nature methods · 2011Article
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
RNA interference (RNAi) is a widely used method for analysis of gene function in tissue culture cells. However, to date there has been no reliable method for testing the specificity of any particular RNAi experiment. The ideal experiment is to rescue the phenotype by expression of the target gene in a form refractory to RNAi. The transgene should be expressed at physiological levels and with its different splice variants. Here, we demonstrate that expression of murine bacterial artificial chromosomes in human cells provides a reliable method to create RNAi-resistant transgenes. This strategy should be applicable to all eukaryotes and should therefore be a standard technology for confirming the specificity of RNAi. We show that this technique can be extended to allow the creation of tagged transgenes, expressed at physiological levels, for the further study of gene function.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.