ArticleAnalytical biochemistry2011
A primer-free method that selects high-affinity single-stranded DNA aptamers using thermostable RNA ligase.
Article in Analytical biochemistry, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 22 citations in OpenAlex.
- Aptamers in Virology-A Consolidated Review of the Most Recent Advancements in Diagnosis and Therapy.Pharmaceutics · 2021Review
- A New Design for the Fixed Primer Regions in an Oligonucleotide Library for SELEX Aptamer Screening.Frontiers in chemistry · 2020Article
- Inside the Black Box: What Makes SELEX Better?Molecules (Basel, Switzerland) · 2019Review
- Lysozyme Aptamer-Functionalized Magnetic Nanoparticles for the Purification of Lysozyme from Chicken Egg White.Foods (Basel, Switzerland) · 2019Article
- Key Aspects of Nucleic Acid Library Design for in Vitro Selection.International journal of molecular sciences · 2018Review
- Aptamer Cell-Based Selection: Overview and Advances.Biomedicines · 2017Review
- Selection of 2'-deoxy-2'-fluoroarabinonucleotide (FANA) aptamers that bind HIV-1 reverse transcriptase with picomolar affinity.Nucleic acids research · 2015Article
- Aptamer Selection Technology and Recent Advances.Molecular therapy. Nucleic acids · 2015Review
- New Technologies Provide Quantum Changes in the Scale, Speed, and Success of SELEX Methods and Aptamer Characterization.Molecular therapy. Nucleic acids · 2014Article
- DNA aptamers to human immunodeficiency virus reverse transcriptase selected by a primer-free SELEX method: characterization and comparison with other aptamers.Nucleic acid therapeutics · 2012Article
- RNA aptamer evolution: two decades of SELEction.International journal of molecular sciences · 2011Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
This article describes a method for selecting single-stranded DNA (ssDNA) molecules that bind with high-affinity aptamers to specific target proteins. This SELEX (systematic evolution of ligands by exponential enrichment) method is similar to other "primer-free" approaches where the random sequence ssDNA starting pool has no fixed sequences at the 5' and 3' termini. Therefore, there are no predetermined sequences that could bias selection. Like other SELEX methods, repeated cycles (typically 5-15) of selection and then amplification and reselection are used. The method differs from other primer-free approaches in that the key step for regenerating new material for subsequent rounds is ligation of the selected ssDNA to a defined sequence oligonucleotide using thermostable RNA ligase. Under specific conditions, this ligase ligated 30-nt random sequence ssDNA (5'-N(30)-3') to a specified 20-nt ssDNA with approximately 50% efficiency. Efficiency was improved to approximately 90% by the addition of a single T residue to the 3' end (5'-N(29)T-3'). High efficiency in this step is critical, especially early in the procedure because any selected material that is not ligated is lost. In this study, human immunodeficiency virus reverse transcriptase was used as the target protein, but the method could be applied to essentially any protein.
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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.