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Here the mechanisms of Cas9 specificity, and strategies to assess and mitigate unwanted effects of Cas9 editing are reviewed; covering guide‐RNA design, RNA modifications, Cas9 modifications, control of Cas9 activity; computational prediction for off‐targets, and experimental methods for detecting Cas9 cleavage. Although recognition of the prevalence of on‐ and off‐target effects of Cas9 editing has increased in recent years, broader uptake across the gene editing community will be important in determining the specificity of Cas9 across diverse applications and organisms.</jats:p>","journal":"BioEssays","year":2020,"id":590838,"datarank":0.8057994397864243,"base_score":3.091042453358316,"endowment":3.091042453358316,"self_citation_contribution":0.4636563680037475,"citation_network_contribution":0.3421430717826769,"self_endowment_contribution":0.4636563680037475,"citer_contribution":0.3421430717826769,"corpus_percentile":null,"corpus_rank":null,"citation_count":21,"citer_count":20,"citers_with_citation_signal":17,"citers_with_endowment":17,"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":1376007,"name":"Lora Starrs","orcid":"0000-0001-5771-4969","position":1,"is_corresponding":false},{"id":285701,"name":"Gaétan Burgio","orcid":"0000-0002-7434-926X","position":2,"is_corresponding":false},{"id":1376006,"name":"Anthony Newman","orcid":"0009-0001-6093-0010","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Cas9 Cuts and Consequences; Detecting, Predicting, and Mitigating CRISPR/Cas9 On‐ and Off‐Target Damage","abstract":"<jats:title>Abstract</jats:title><jats:p>Large deletions and genomic re‐arrangements are increasingly recognized as common products of double‐strand break repair at Clustered Regularly Interspaced, Short Palindromic Repeats ‐ CRISPR associated protein 9 (CRISPR/Cas9) on‐target sites. Together with well‐known off‐target editing products from Cas9 target misrecognition, these are important limitations, that need to be addressed. Rigorous assessment of Cas9‐editing is necessary to ensure validity of observed phenotypes in Cas9‐edited cell‐lines and model organisms. Here the mechanisms of Cas9 specificity, and strategies to assess and mitigate unwanted effects of Cas9 editing are reviewed; covering guide‐RNA design, RNA modifications, Cas9 modifications, control of Cas9 activity; computational prediction for off‐targets, and experimental methods for detecting Cas9 cleavage. Although recognition of the prevalence of on‐ and off‐target effects of Cas9 editing has increased in recent years, broader uptake across the gene editing community will be important in determining the specificity of Cas9 across diverse applications and organisms.</jats:p>","is_dataset_classified":null,"base_score":3.091042453358316,"endowment":3.091042453358316,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"32643177","pmcid":null,"openalex_id":"https://openalex.org/W3040709352","authors":[],"funders":[{"funder_name":"National Natural Science Foundation of China","grant_id":"81772214","title":null},{"funder_name":"Australian Research Council","grant_id":"DP180101494","title":"Discovery Projects - Grant ID: DP180101494"},{"funder_name":"National Health and Medical Research Council","grant_id":"APP1143008","title":null},{"funder_name":"Australian Government Research Training Program Scholarship","grant_id":"","title":null},{"funder_name":"Phenomics Australian","grant_id":"","title":null},{"funder_name":"National Collaborative Research Infrastructure Strategy (NCRIS)","grant_id":"","title":null},{"funder_name":"National Collaborative Research Infrastructure","grant_id":"","title":null},{"funder_name":"National Collaborative Research Infrastructure Strategy (NCRIS)","grant_id":"","title":null},{"funder_name":"Phenomics Australian","grant_id":"","title":null},{"funder_name":"Australian Government Research Training Program Scholarship","grant_id":"","title":null},{"funder_name":"National Collaborative Research Infrastructure","grant_id":"","title":null}],"total_grants":11,"fwci":0.895,"citation_percentile":0.72490848,"influential_citations":0,"citation_trend":[{"year":2020,"count":1},{"year":2021,"count":3},{"year":2022,"count":2},{"year":2023,"count":5},{"year":2024,"count":4},{"year":2025,"count":5},{"year":2026,"count":1}],"oa_status":"closed","license":"Wiley Online Library User Agreement","oa_locations":[{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fbies.202000047","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1002/bies.202000047","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/full-xml/10.1002/bies.202000047","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/am-pdf/10.1002/bies.202000047","host_type":"publisher"},{"url":"https://doi.org/10.1002/bies.202000047","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/32643177","host_type":"repository"},{"url":"https://rss.onlinelibrary.wiley.com/doi/am-pdf/10.1002/bies.202000047","host_type":""},{"url":"https://dx.doi.org/10.1002/bies.202000047","host_type":""}],"fields_of_study":["CRISPR and Genetic Engineering","RNA regulation and disease","Retinal Development and Disorders","0301 basic medicine","0303 health sciences","03 medical and health sciences","CRISPR-Cas Systems","Gene Editing","Genome","Genomics","Humans","RNA, Guide, CRISPR-Cas Systems"],"mesh_terms":["Gene Editing","RNA, Guide, CRISPR-Cas Systems","Humans","Genome","RNA, Guide, Kinetoplastida","Genomics","CRISPR-Cas Systems"],"keywords":["CRISPR","Cas9","Genome editing","Computational biology","Biology","Palindrome","Guide RNA","Double strand","RNA","Gene","Genetics","Computer science","DNA repair","Machine Learning","High-throughput Sequencing","Off-target Effects","Crispr/cas9","Double-stranded Breaks","Guide Rna Design","On-target Effects","Gene Editing","Genome","Humans","Genomics","CRISPR-Cas Systems","RNA, Guide, CRISPR-Cas Systems"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-25T12:36:18.226429Z","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":[]}