{"doi":"10.1016/j.bios.2019.111361","title":"Use of anti-CRISPR protein AcrIIA4 as a capture ligand for CRISPR/Cas9 detection","abstract":null,"journal":"Biosensors and Bioelectronics","year":2019,"id":603705,"datarank":0.5244761342199721,"base_score":3.4965075614664802,"endowment":3.4965075614664802,"self_citation_contribution":0.5244761342199721,"citation_network_contribution":0.0,"self_endowment_contribution":0.5244761342199721,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":32,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":733622,"name":"Kyle J. Seamon","orcid":"0009-0005-5028-3752","position":1,"is_corresponding":false},{"id":494443,"name":"Edwin A. Saada","orcid":"0000-0003-3699-1849","position":2,"is_corresponding":false},{"id":472016,"name":"Joshua D. Podlevsky","orcid":"0000-0001-5938-2478","position":3,"is_corresponding":false},{"id":579479,"name":"Steven S. Branda","orcid":"0000-0001-8763-1013","position":4,"is_corresponding":false},{"id":1548705,"name":"Jerilyn A. Timlin","orcid":null,"position":5,"is_corresponding":false},{"id":1548708,"name":"Jason C. Harper","orcid":null,"position":6,"is_corresponding":false},{"id":1548699,"name":"Robert K. Johnston","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Use of anti-CRISPR protein AcrIIA4 as a capture ligand for CRISPR/Cas9 detection","abstract":"The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) ribonucleoprotein (RNP) complex is an RNA-guided DNA-nuclease that is part of the bacterial adaptive immune system. CRISPR/Cas9 RNP has been adapted for targeted genome editing within cells and whole organisms with new applications vastly outpacing detection and quantification of gene-editing reagents. Detection of the CRISPR/Cas9 RNP within biological samples is critical for assessing gene-editing reagent delivery efficiency, retention, persistence, and distribution within living organisms. Conventional detection methods are effective, yet the expense and lack of scalability for antibody-based affinity reagents limit these techniques for clinical and/or field settings. This necessitates the development of low cost, scalable CRISPR/Cas9 RNP affinity reagents as alternatives or augments to antibodies. Herein, we report the development of the Streptococcus pyogenes anti-CRISPR/Cas9 protein, AcrIIA4, as a novel affinity reagent. An engineered cysteine linker enables covalent immobilization of AcrIIA4 onto glassy carbon electrodes functionalized via aryl diazonium chemistry for detection of CRISPR/Cas9 RNP by electrochemical, fluorescent, and colorimetric methods. Electrochemical measurements achieve a detection of 280 pM RNP in reaction buffer and 8 nM RNP in biologically representative conditions. Our results demonstrate the ability of anti-CRISPR proteins to serve as robust, specific, flexible, and economical recognition elements in biosensing/quantification devices for CRISPR/Cas9 RNP.","is_dataset_classified":null,"base_score":3.4965075614664802,"endowment":3.4965075614664802,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"31207570","pmcid":null,"openalex_id":"https://openalex.org/W2947638201","authors":[],"funders":[{"funder_name":"Sandia National Laboratories","grant_id":"","title":null},{"funder_name":"Defense Advanced Research Projects Agency","grant_id":"","title":null}],"total_grants":2,"fwci":1.1384,"citation_percentile":0.77312577,"influential_citations":0,"citation_trend":[{"year":2019,"count":2},{"year":2020,"count":5},{"year":2021,"count":6},{"year":2022,"count":2},{"year":2023,"count":6},{"year":2024,"count":4},{"year":2025,"count":2},{"year":2026,"count":5}],"oa_status":"green","license":"http://www.elsevier.com/open-access/userlicense/1.0/","oa_locations":[{"url":"https://www.osti.gov/biblio/1559508","host_type":"repository"},{"url":"https://www.osti.gov/biblio/1559508","host_type":"repository"},{"url":"https://api.elsevier.com/content/article/PII:S0956566319304403?httpAccept=text/xml","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S0956566319304403?httpAccept=text/plain","host_type":"publisher"},{"url":"https://doi.org/10.1016/j.bios.2019.111361","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/31207570","host_type":"repository"},{"url":"https://www.osti.gov/biblio/1778458","host_type":"repository"}],"fields_of_study":["CRISPR and Genetic Engineering","Cytomegalovirus and herpesvirus research","Viral Infections and Immunology Research","Bacterial Proteins","Bacteriophages","Biosensing Techniques","CRISPR-Associated Protein 9","CRISPR-Cas Systems","Immobilized Proteins","Ligands","Models, Molecular","Streptococcus pyogenes","Viral Proteins"],"mesh_terms":["CRISPR-Associated Protein 9","Bacterial Proteins","Bacteriophages","Ligands","Models, Molecular","Streptococcus pyogenes","Viral Proteins","Biosensing Techniques","Immobilized Proteins","CRISPR-Cas Systems"],"keywords":["CRISPR","Cas9","Trans-activating crRNA","Ribonucleoprotein","Nuclease","Genome editing","Computational biology","Guide RNA","Chemistry","DNA","RNA","Biology","Gene","Biochemistry","Electrochemistry","Affinity Reagent","Crispr/cas9","Anti-crispr Protein"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-29T22:32:22.866258Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}