ArticleNucleic acids research2022
Directed evolution of orthogonal RNA-RBP pairs through library-vs-library in vitro selection.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed, 16 citations in OpenAlex.
- Decoding RNA-protein interactions using high-throughput methods.RNA biology · 2026Review
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- Systematic Evaluation of Different Ribonucleoprotein Complexes as Posttranscriptional Biosensors in Cell-Free TX-TL Systems.ACS synthetic biology · 2026Article
- Bioluminescent Probes for Multiplexed RNA Imaging.Journal of the American Chemical Society · 2026Article
- Protein-inducible ribosomal frameshifting enables programmable translational control for genetic circuit design in Escherichia coli.Journal of biological engineering · 2026Article
- Expanding the σ54-dependent transcription process with orthogonal designs.Nucleic acids research · 2025Article
- Development of multiplexed orthogonal base editor (MOBE) systems.Nature biotechnology · 2025Article
- Structural insights into lab-coevolved RNA-RBP pairs and applications of synthetic riboswitches in cell-free system.Nucleic acids research · 2025Article
- Article
- Accelerating Genetic Sensor Development, Scale-up, and Deployment Using Synthetic Biology.Biodesign research · 2024Article
- Sensing intracellular signatures with synthetic mRNAs.RNA biology · 2023Review
- Methods for the directed evolution of biomolecular interactions.Trends in biochemical sciences · 2022Review
- Protein-Based Systems for Translational Regulation of Synthetic mRNAs in Mammalian Cells.Life (Basel, Switzerland) · 2021Review
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2 authors at 1 institution in 1 country.
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Abstract
RNA-binding proteins (RBPs) and their RNA ligands play many critical roles in gene regulation and RNA processing in cells. They are also useful for various applications in cell biology and synthetic biology. However, re-engineering novel and orthogonal RNA-RBP pairs from natural components remains challenging while such synthetic RNA-RBP pairs could significantly expand the RNA-RBP toolbox for various applications. Here, we report a novel library-vs-library in vitro selection strategy based on Phage Display coupled with Systematic Evolution of Ligands by EXponential enrichment (PD-SELEX). Starting with pools of 1.1 × 1012 unique RNA sequences and 4.0 × 108 unique phage-displayed L7Ae-scaffold (LS) proteins, we selected RNA-RBP complexes through a two-step affinity purification process. After six rounds of library-vs-library selection, the selected RNAs and LS proteins were analyzed by next-generation sequencing (NGS). Further deconvolution of the enriched RNA and LS protein sequences revealed two synthetic and orthogonal RNA-RBP pairs that exhibit picomolar affinity and >4000-fold selectivity.
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