ArticleThe Biochemical journal2002
Isoform-specific knockdown and expression of adaptor protein ShcA using small interfering RNA.
Article in The Biochemical journal, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 88 citations in OpenAlex.
- P46Shc Inhibits Mitochondrial ACAA2 Thiolase, Exacerbating Mitochondrial Injury and Inflammation in Aging Livers.The American journal of pathology · 2025Article
- Distribution of the p66Shc Adaptor Protein Among Mitochondrial and Mitochondria-Associated Membranes Fractions in Normal and Oxidative Stress Conditions.International journal of molecular sciences · 2024Article
- Article
- Isoform specific knockdown of the ETS transcription factor Pointed inmicroPublication biology · 2023Article
- Bioinformatics Pipeline to Analyze lncRNA Arrays.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Prognostic value and prospective molecular mechanism of miR-100-5p in hepatocellular carcinoma: A comprehensive study based on 1,258 samples.Oncology letters · 2019Article
- Gene-level differential analysis at transcript-level resolution.Genome biology · 2018Article
- Loss of the adaptor protein ShcA in endothelial cells protects against monocyte macrophage adhesion, LDL-oxydation, and atherosclerotic lesion formation.Scientific reports · 2018Article
- p46Shc Inhibits Thiolase and Lipid Oxidation in Mitochondria.The Journal of biological chemistry · 2016Article
- Non-canonical dynamic mechanisms of interaction between the p66Shc protein and Met receptor.The Biochemical journal · 2016Article
- ShcA Protects against Epithelial-Mesenchymal Transition through Compartmentalized Inhibition of TGF-β-Induced Smad Activation.PLoS biology · 2015Article
- Diversity in TAF proteomics: consequences for cellular differentiation and migration.International journal of molecular sciences · 2014Review
- Article
- The adaptor proteins p66Shc and Grb2 regulate the activation of the GTPases ARF1 and ARF6 in invasive breast cancer cells.The Journal of biological chemistry · 2014Article
- The adaptor protein p66Shc inhibits mTOR-dependent anabolic metabolism.Science signaling · 2014Article
- The p66Shc adapter protein regulates the morphogenesis and epithelial maturation of fetal mouse lungs.American journal of physiology. Lung cellular and molecular physiology · 2014Article
- TNFα signals via p66(Shc) to induce E-Selectin, promote leukocyte transmigration and enhance permeability in human endothelial cells.PloS one · 2013Article
- Host gene targets for novel influenza therapies elucidated by high-throughput RNA interference screens.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2012Review
- The Shc locus regulates insulin signaling and adiposity in mammals.Aging cell · 2011Article
- Decreased superoxide production in macrophages of long-lived p66Shc knock-out mice.The Journal of biological chemistry · 2010Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Many eukaryotic genes are expressed as multiple isoforms through the differential utilization of transcription/translation initiation sites or alternative splicing. The conventional approach for studying individual isoforms in a clean background (i.e. without the influence of other isoforms) has been to express them in cells or whole organisms in which the target gene has been deleted; this is time-consuming. Recently an efficient post-transcriptional gene-silencing method has been reported that employs a small interfering double-stranded RNA (siRNA). On the basis of this method we report a rapid alternative approach for isoform-specific gene expression. We show how the adaptor protein ShcA can be suppressed and expressed in an isoform-specific manner in a human cell line. ShcA exists in three isoforms, namely p66, p52 and p46, which differ only in their N-terminal regions and are derived from two different transcripts, namely p66 and p52/p46 mRNAs. An siRNA with a sequence shared by the two transcripts suppressed all of them. However, another siRNA whose sequence was present only in p66 mRNA suppressed only the p66 isoform, suggesting that the siRNA signal did not propagate to other regions of the target mRNA. The expression of individual isoforms was achieved by first down-regulating all isoforms by the common siRNA and then transfecting with an expression vector for each isoform that harboured silent mutations at the site corresponding to the siRNA. This allowed functional analysis of individual ShcA isoforms and may be more generally applicable for studying genes encoding multiple proteins.
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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.