ArticlePloS one2013
AAscan, PCRdesign and MutantChecker: a suite of programs for primer design and sequence analysis for high-throughput scanning mutagenesis.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- MAXTIA: A high-throughput platform for rapid functional epitope mapping by kinetic screening of mutant libraries.Protein science : a publication of the Protein Society · 2026Article
- Phosphorylation site topology governs the functional dynamics of arrestin recruitment to GPCRs.Research square · 2026Article
- Article
- Enriching stabilizing mutations through automated analysis of molecular dynamics simulations using BoostMut.Protein science : a publication of the Protein Society · 2025Article
- FASTIA: A rapid and accessible platform for protein variant interaction analysis demonstrated with a single-domain antibody.Protein science : a publication of the Protein Society · 2025Article
- Designing Michaelases: exploration of novel protein scaffolds for iminium biocatalysis.Faraday discussions · 2024Article
- Spectroelectrochemistry for determination of the redox potential in heme enzymes: Dye-decolorizing peroxidases.BBA advances · 2024Article
- Molecular determinants of ligand efficacy and potency in GPCR signaling.Science (New York, N.Y.) · 2023Article
- Structural details of a Class B GPCR-arrestin complex revealed by genetically encoded crosslinkers in living cells.Nature communications · 2023Article
- The pocketome of G-protein-coupled receptors reveals previously untargeted allosteric sites.Nature communications · 2022Article
- Folding and Evolution of a Repeat Protein on the Ribosome.Frontiers in molecular biosciences · 2022Article
- Residue-level determinants of RGS R4 subfamily GAP activity and specificity towards the GCellular and molecular life sciences : CMLS · 2021Article
- Exploring GPCR-arrestin interfaces with genetically encoded crosslinkers.EMBO reports · 2020Article
- Capturing Peptide-GPCR Interactions and Their Dynamics.Molecules (Basel, Switzerland) · 2020Review
- High-throughput Site-directed Scanning Mutagenesis Using a Two-fragment PCR Approach.Bio-protocol · 2020Article
- Evolving a Peptide: Library Platforms and Diversification Strategies.International journal of molecular sciences · 2019Review
- Stabilization of cyclohexanone monooxygenase by computational and experimental library design.Biotechnology and bioengineering · 2019Article
- Article
- Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation.International journal of molecular sciences · 2019Review
- Arrestin-1 engineering facilitates complex stabilization with native rhodopsin.Scientific reports · 2019Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Scanning mutagenesis is a powerful protein engineering technique used to study protein structure-function relationship, map binding sites and design more stable proteins or proteins with altered properties. One of the time-consuming tasks encountered in application of this technique is the design of primers for site-directed mutagenesis. Here we present an open-source multi-platform software AAscan developed to design primers for this task according to a set of empirical rules such as melting temperature, overall length, length of overlap regions, and presence of GC clamps at the 3' end, for any desired substitution. We also describe additional software tools which are used to analyse a large number of sequencing results for the presence of desired mutations, as well as related software to design primers for ligation independent cloning. We have used AAscan software to design primers to make over 700 mutants, with a success rate of over 80%. We hope that the open-source nature of our software and ready availability of freeware tools used for its development will facilitate its adaptation and further development. The software is distributed under GPLv3 licence and is available at http://www.psi.ch/lbr/aascan.
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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.