{"doi":"10.3390/ijms20153719","title":"Strategies to Increase On-Target and Reduce Off-Target Effects of the CRISPR/Cas9 System in Plants","abstract":"<jats:p>The CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeat-associated protein 9) is a powerful genome-editing tool in animals, plants, and humans. This system has some advantages, such as a high on-target mutation rate (targeting efficiency), less cost, simplicity, and high-efficiency multiplex loci editing, over conventional genome editing tools, including meganucleases, transcription activator-like effector nucleases (TALENs), and zinc finger nucleases (ZFNs). One of the crucial shortcomings of this system is unwanted mutations at off-target sites. We summarize and discuss different approaches, such as dCas9 and Cas9 paired nickase, to decrease the off-target effects in plants. According to studies, the most effective method to reduce unintended mutations is the use of ligand-dependent ribozymes called aptazymes. The single guide RNA (sgRNA)/ligand-dependent aptazyme strategy has helped researchers avoid unwanted mutations in human cells and can be used in plants as an alternative method to dramatically decrease the frequency of off-target mutations. We hope our concept provides a new, simple, and fast gene transformation and genome-editing approach, with advantages including reduced time and energy consumption, the avoidance of unwanted mutations, increased frequency of on-target changes, and no need for external forces or expensive equipment.</jats:p>","journal":"International Journal of Molecular Sciences","year":2019,"id":612201,"datarank":2.536692169762159,"base_score":4.709530201312334,"endowment":4.709530201312334,"self_citation_contribution":0.7064295301968502,"citation_network_contribution":1.8302626395653088,"self_endowment_contribution":0.7064295301968502,"citer_contribution":1.8302626395653088,"corpus_percentile":null,"corpus_rank":null,"citation_count":110,"citer_count":106,"citers_with_citation_signal":87,"citers_with_endowment":87,"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":1576158,"name":"Ali Movahedi","orcid":"0000-0001-5062-504X","position":1,"is_corresponding":false},{"id":1349139,"name":"Hui Wei","orcid":"0000-0002-2462-8180","position":2,"is_corresponding":false},{"id":968465,"name":"Dawei Li","orcid":"0000-0002-0840-0602","position":3,"is_corresponding":false},{"id":1576159,"name":"Yasin Orooji","orcid":"0000-0001-6940-5047","position":4,"is_corresponding":false},{"id":1576160,"name":"Honghua Ruan","orcid":null,"position":5,"is_corresponding":false},{"id":1576161,"name":"Qiang Zhuge","orcid":null,"position":6,"is_corresponding":false},{"id":1576157,"name":"Zahra Hajiahmadi","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Strategies to Increase On-Target and Reduce Off-Target Effects of the CRISPR/Cas9 System in Plants.","abstract":"The CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeat-associated protein 9) is a powerful genome-editing tool in animals, plants, and humans. This system has some advantages, such as a high on-target mutation rate (targeting efficiency), less cost, simplicity, and high-efficiency multiplex loci editing, over conventional genome editing tools, including meganucleases, transcription activator-like effector nucleases (TALENs), and zinc finger nucleases (ZFNs). One of the crucial shortcomings of this system is unwanted mutations at off-target sites. We summarize and discuss different approaches, such as dCas9 and Cas9 paired nickase, to decrease the off-target effects in plants. According to studies, the most effective method to reduce unintended mutations is the use of ligand-dependent ribozymes called aptazymes. The single guide RNA (sgRNA)/ligand-dependent aptazyme strategy has helped researchers avoid unwanted mutations in human cells and can be used in plants as an alternative method to dramatically decrease the frequency of off-target mutations. We hope our concept provides a new, simple, and fast gene transformation and genome-editing approach, with advantages including reduced time and energy consumption, the avoidance of unwanted mutations, increased frequency of on-target changes, and no need for external forces or expensive equipment.","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"31366028","pmcid":"PMC6696359","openalex_id":null,"authors":[],"funders":[{"funder_name":"Nanjing Forestry University","grant_id":"163108059","title":null}],"total_grants":1,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://www.mdpi.com/1422-0067/20/15/3719/pdf?version=1564468112","host_type":"publisher"},{"url":"https://doaj.org/article/e83022ad1ca9434d86983a090d723254","host_type":"repository"},{"url":"http://dx.doi.org/10.3390/ijms20153719","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/6696359","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC6696359","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC6696359?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":[],"mesh_terms":["Targeted Gene Repair","CRISPR-Cas Systems","Plant Breeding","Gene Editing","Magnoliopsida","RNA, Guide, CRISPR-Cas Systems"],"keywords":["Crispr","Off-target Effect","Cas9","Aptazyme","On-target Mutation"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"doi"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-02T02:08:31.711760Z","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":[]}