ArticleProceedings of the National Academy of Sciences of the United States of America2001
T7 phage display: a novel genetic selection system for cloning RNA-binding proteins from cDNA libraries.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2001. 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.
- Decoding RNA-protein interactions using high-throughput methods.RNA biology · 2026Review
- Article
- Enhancement of plasma kallikrein specificity of antitrypsin variants identified by phage display and partial reversion.BMC biotechnology · 2025Article
- BacScan: a novel genome-wide strategy for uncovering broadly immunogenic proteins in bacteria.Frontiers in immunology · 2024Article
- Review
- Sensing intracellular signatures with synthetic mRNAs.RNA biology · 2023Review
- Glycan-Adhering Lectins and Experimental Evaluation of a Lectin FimH Inhibitor in EnterohemorrhagicInternational journal of molecular sciences · 2022Article
- The Screening of Therapeutic Peptides for Anti-Inflammation through Phage Display Technology.International journal of molecular sciences · 2022Review
- Analyzing the Androgen Receptor Interactome in Prostate Cancer: Implications for Therapeutic Intervention.Cells · 2022Review
- Screening of TNFR1 Binding Peptides fromToxins · 2022Article
- T7 Phage as an Emerging Nanobiomaterial with Genetically Tunable Target Specificity.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2022Review
- Directed evolution of orthogonal RNA-RBP pairs through library-vs-library in vitro selection.Nucleic acids research · 2022Article
- Heat adaptation of phage T7 under an extended genetic code.Virus evolution · 2021Article
- A Homeobox Transcription Factor Scarecrow (SCRO) Negatively Regulates Pdf Neuropeptide Expression through Binding an Identified cis-Acting Element in Drosophila melanogaster.Molecular neurobiology · 2020Article
- A novel approach for T7 bacteriophage genome integration of exogenous DNA.Journal of biological engineering · 2020Article
- Review
- Plant/Bacterial Virus-Based Drug Discovery, Drug Delivery, and Therapeutics.Pharmaceutics · 2019Review
- Bacteriophage T4 nanoparticles for vaccine delivery against infectious diseases.Advanced drug delivery reviews · 2019Review
- Genetic Engineering of Bacteriophages Against Infectious Diseases.Frontiers in microbiology · 2019Review
- Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing.Journal of visualized experiments : JoVE · 2018Article
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2 authors.
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Abstract
RNA-binding proteins are central to posttranscriptional gene regulation and play an important role in a number of major human diseases. Cloning such proteins is a crucial but often difficult step in elucidating the biological function of RNA regulatory elements. To make it easier to clone proteins that specifically bind RNA elements of interest, we have developed a rapid and broadly applicable in vitro genetic selection method based on T7 phage display. Using hairpin II of U1 small nuclear RNA (U1hpII) or the 3' stem loop of histone mRNA as bait, we could selectively amplify T7 phage that display either the spliceosomal protein U1A or the histone stem loop-binding protein from a lung cDNA phage library containing more than 10(7) independent clones. The use of U1hpII mutants with various affinities for U1A revealed that this method allows the selection even of proteins that bind their cognate RNA targets with relatively weak affinities (K(d) as high as the micromolar range). Experiments with a mixture of recombinant phage displaying U1A or the closely related protein U2B" demonstrated that addition of a competitor RNA can suppress selection of a protein with a higher affinity for a given RNA target, thereby allowing the preferential amplification of a lower affinity protein. Together, these findings suggest that T7 phage display can be used to rapidly and selectively clone virtually any protein that binds a known RNA regulatory element, including those that bind with low affinity or that must compete for binding with other proteins.
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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.