ArticleRNA (New York, N.Y.)2015
Using droplet-based microfluidics to improve the catalytic properties of RNA under multiple-turnover conditions.
Article in RNA (New York, N.Y.), 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
34 citing papers in PubMed, 77 citations in OpenAlex.
- Extending the Potential of Digital Droplet PCR by Expressing Amplified Genes and Assaying Their Phenotype.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Evolution Driven Microscale Combinatorial Chemistry in Intracellular Mimicking Droplets to Engineer Thermostable RNA for Cellular Imaging.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- From specialization to broad adoption: Key trends in droplet microfluidic innovations enhancing accessibility to non-experts.Biomicrofluidics · 2025Article
- Cross-chiral exponential amplification of an RNA enzyme.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Structure-based insights into fluorogenic RNA aptamers.Acta biochimica et biophysica Sinica · 2024Review
- Droplet Microfluidics for High-Throughput Screening and Directed Evolution of Biomolecules.Micromachines · 2024Review
- Ultrahigh-Throughput Enzyme Engineering and Discovery inChemical reviews · 2023Review
- Design and construction of a microfluidics workstation for high-throughput multi-wavelength fluorescence and transmittance activated droplet analysis and sorting.Nature protocols · 2023Review
- Cell-Free Production Systems in Droplet Microfluidics.Advances in biochemical engineering/biotechnology · 2023Review
- Aptamers as Functional Modules for DNA Nanostructures.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Pushing the Limits of Nucleic Acid Function.Chemistry (Weinheim an der Bergstrasse, Germany) · 2022Review
- High-throughput screening of cell-free riboswitches by fluorescence-activated droplet sorting.Nucleic acids research · 2022Article
- Directed Evolution in Drops: Molecular Aspects and Applications.ACS synthetic biology · 2021Review
- Digital Microfluidics Chips for the Execution and Real-Time Monitoring of Multiple Ribozymatic Cleavage Reactions.ACS omega · 2021Article
- Water/Oil Emulsions with Controlled Droplet Sizes forACS omega · 2021Article
- μIVC-Seq: A Method for Ultrahigh-Throughput Development and Functional Characterization of Small RNAs.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Microfluidic droplet generation based on non-embedded co-flow-focusing using 3D printed nozzle.Scientific reports · 2020Article
- The nature of the purine at position 34 in tRNAs of 4-codon boxes is correlated with nucleotides at positions 32 and 38 to maintain decoding fidelity.Nucleic acids research · 2020Article
- A dimerization-based fluorogenic dye-aptamer module for RNA imaging in live cells.Nature chemical biology · 2020Article
- High-throughput droplet-based microfluidics for directed evolution of enzymes.Electrophoresis · 2019Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In vitro evolution methodologies are powerful approaches to identify RNA with new functionalities. While Systematic Evolution of Ligands by Exponential enrichment (SELEX) is an efficient approach to generate new RNA aptamers, it is less suited for the isolation of efficient ribozymes as it does not select directly for the catalysis. In vitro compartmentalization (IVC) in aqueous droplets in emulsions allows catalytic RNAs to be selected under multiple-turnover conditions but suffers severe limitations that can be overcome using the droplet-based microfluidics workflow described in this paper. Using microfluidics, millions of genes in a library can be individually compartmentalized in highly monodisperse aqueous droplets and serial operations performed on them. This allows the different steps of the evolution process (gene amplification, transcription, and phenotypic assay) to be uncoupled, making the method highly flexible, applicable to the selection and evolution of a variety of RNAs, and easily adaptable for evolution of DNA or proteins. To demonstrate the method, we performed cycles of random mutagenesis and selection to evolve the X-motif, a ribozyme which, like many ribozymes selected using SELEX, has limited multiple-turnover activity. This led to the selection of variants, likely to be the optimal ribozymes that can be generated using point mutagenesis alone, with a turnover number under multiple-turnover conditions, k(ss) cat, ∼ 28-fold higher than the original X-motif, primarily due to an increase in the rate of product release, the rate-limiting step in the multiple-turnover reaction.
Indexed as
Identifiers
What OpenQuestion holds
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.