{"doi":"10.3389/fmicb.2021.686715","title":"Functional Identification of the Xanthomonas oryzae pv. oryzae Type I-C CRISPR-Cas System and Its Potential in Gene Editing Application","abstract":"<jats:p>The type I clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system is one of five adaptive immune systems and exists widely in bacteria and archaea. In this study, we showed that <jats:italic>Xanthomonas oryzae</jats:italic> pv. <jats:italic>oryzae</jats:italic> (<jats:italic>Xoo</jats:italic>) possesses a functional CRISPR system by engineering constructs mimicking its CRISPR cassette. CRISPR array analysis showed that the TTC at the 5′-end of the target sequence is a functional protospacer-adjacent motif (PAM) of CRISPR. Guide RNA (gRNA) deletion analysis identified a minimum of 27-bp spacer that was required to ensure successful self-target killing in PXO99<jats:sup>A</jats:sup> strain. Mutants with deletion of individual <jats:italic>Cas</jats:italic> genes were constructed to analyze the effects of Cas proteins on mature CRISPR RNA (crRNA), processing intermediates and DNA interference. Results showed that depleting each of the three genes, <jats:italic>cas5d</jats:italic>, <jats:italic>csd1</jats:italic>, and <jats:italic>csd2</jats:italic> inactivated the pre-crRNA processing, whereas inactivation of <jats:italic>cas3</jats:italic> impaired in processing pre-crRNA. Furthermore, the <jats:italic>Xoo</jats:italic> CRISPR/Cas system was functional in <jats:italic>Pseudomonas syringae</jats:italic> pv. <jats:italic>tomato</jats:italic>. Collectively, our results would contribute to the functional study of CRISPR/Cas system of <jats:italic>Xoo</jats:italic>, and also provide a new vision on the use of bacterial endogenous systems as a convenient tool for gene editing.</jats:p>","journal":"Frontiers in Microbiology","year":2021,"id":45637,"datarank":0.5865597758775472,"base_score":2.4849066497880004,"endowment":2.4849066497880004,"self_citation_contribution":0.37273599746820013,"citation_network_contribution":0.21382377840934705,"self_endowment_contribution":0.37273599746820013,"citer_contribution":0.21382377840934705,"corpus_percentile":null,"corpus_rank":null,"citation_count":11,"citer_count":11,"citers_with_citation_signal":8,"citers_with_endowment":8,"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":212927,"name":"Siwei Wang","orcid":null,"position":1,"is_corresponding":false},{"id":212928,"name":"Juying Long","orcid":null,"position":2,"is_corresponding":false},{"id":212929,"name":"Zhuoyue Chen","orcid":null,"position":3,"is_corresponding":false},{"id":111521,"name":"Bing Yang","orcid":null,"position":4,"is_corresponding":false},{"id":212930,"name":"Fei Lin","orcid":null,"position":5,"is_corresponding":false},{"id":212926,"name":"Qibing Liu","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":2.4849066497880004,"endowment":2.4849066497880004,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"34475857","pmcid":"PMC8406640","openalex_id":"https://openalex.org/W3195673132","authors":[],"funders":[],"total_grants":0,"fwci":0.5699,"citation_percentile":0.64745614,"influential_citations":0,"citation_trend":[{"year":2022,"count":3},{"year":2023,"count":2},{"year":2024,"count":2},{"year":2025,"count":4}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://www.frontiersin.org/articles/10.3389/fmicb.2021.686715/pdf","host_type":"journal"},{"url":"https://www.frontiersin.org/articles/10.3389/fmicb.2021.686715/pdf","host_type":"GOLD"},{"url":"https://www.frontiersin.org/articles/10.3389/fmicb.2021.686715/pdf","host_type":"publisher"},{"url":"https://www.frontiersin.org/articles/10.3389/fmicb.2021.686715/full","host_type":"publisher"},{"url":"https://doi.org/10.3389/fmicb.2021.686715","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/34475857","host_type":"repository"},{"url":"https://doaj.org/article/19996ac2d045427abaac240b8d6d4055","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/8406640","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC8406640","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC8406640?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["CRISPR and Genetic Engineering","Aquaculture disease management and microbiota","Vibrio bacteria research studies","Medicine","Biology","Engineering"],"mesh_terms":[],"keywords":["Trans-activating crRNA","CRISPR","Biology","Xanthomonas oryzae","CRISPR interference","Gene","Genetics","Xanthomonas","Computational biology","Cas9","Guide RNA","RNA","Genome Editing","Cas Protein","Xanthomonas Oryzae Pv. Oryzae (Xoo)","Crispr Array","Type I Crispr-Cas Systems"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Life in Land"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-03T23:15:08.312311Z","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":[]}