{"doi":"10.1371/journal.pone.0198714","title":"Highly multiplexed genome engineering using CRISPR/Cas9 gRNA arrays","abstract":null,"journal":"PLOS ONE","year":2018,"id":592609,"datarank":2.607633424060018,"base_score":4.356708826689592,"endowment":4.356708826689592,"self_citation_contribution":0.6535063240034389,"citation_network_contribution":1.9541271000565794,"self_endowment_contribution":0.6535063240034389,"citer_contribution":1.9541271000565794,"corpus_percentile":null,"corpus_rank":null,"citation_count":77,"citer_count":73,"citers_with_citation_signal":57,"citers_with_endowment":57,"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":555935,"name":"Natalie K. Wolf","orcid":"0000-0002-2015-3384","position":1,"is_corresponding":false},{"id":555934,"name":"Walker S. Lahr","orcid":"0000-0002-3288-3843","position":2,"is_corresponding":false},{"id":735643,"name":"Madison T. Weg","orcid":null,"position":3,"is_corresponding":false},{"id":633216,"name":"Mitchell G. Kluesner","orcid":"0000-0001-5828-9204","position":4,"is_corresponding":false},{"id":553486,"name":"Samantha Lee","orcid":"0000-0002-5309-4915","position":5,"is_corresponding":false},{"id":1516493,"name":"Kai Hui","orcid":null,"position":6,"is_corresponding":false},{"id":1516494,"name":"Masano Shiraiwa","orcid":null,"position":7,"is_corresponding":false},{"id":254328,"name":"Beau R. Webber","orcid":"0000-0002-3950-8747","position":8,"is_corresponding":false},{"id":254329,"name":"Branden S. Moriarity","orcid":"0000-0002-3031-3767","position":9,"is_corresponding":false},{"id":1516492,"name":"Morito Kurata","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Highly multiplexed genome engineering using CRISPR/Cas9 gRNA arrays","abstract":"The CRISPR/Cas9 system is an RNA guided nuclease system that evolved as a mechanism of adaptive immunity in bacteria. This system has been adopted for numerous genome engineering applications in research and recently, therapeutics. The CRISPR/Cas9 system has been largely implemented by delivery of Cas9 as protein, RNA, or plasmid along with a chimeric crRNA-tracrRNA guide RNA (gRNA) under the expression of a pol III promoter, such as U6. Using this approach, multiplex genome engineering has been achieved by delivering several U6-gRNA plasmids targeting multiple loci. However, this approach is limited due to the efficiently of delivering multiple plasmids to a single cell at one time. To augment the capability and accessibility of multiplexed genome engineering, we developed an efficient golden gate based method to assemble gRNAs linked by optimal Csy4 ribonuclease sequences to deliver up to 10 gRNAs as a single gRNA array transcript. Here we report the optimal expression of our guide RNA array under a strong pol II promoter. This system can be implemented alongside the myriad of CRISPR applications, allowing users to model complex biological processes requiring numerous gRNAs.","is_dataset_classified":null,"base_score":4.356708826689592,"endowment":4.356708826689592,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"30222773","pmcid":"PMC6141065","openalex_id":"https://openalex.org/W2950422347","authors":[],"funders":[{"funder_name":"National Cancer Institute grant","grant_id":"R03 1R03CA201502-01","title":null},{"funder_name":"NCI NIH HHS","grant_id":"R03 CA201502","title":null},{"funder_name":"Zach Sobiech Osteosarcoma Fund","grant_id":"","title":null},{"funder_name":"The Children’s Cancer Research Fund","grant_id":"","title":null}],"total_grants":4,"fwci":2.3894,"citation_percentile":0.90133555,"influential_citations":0,"citation_trend":[{"year":2018,"count":1},{"year":2019,"count":2},{"year":2020,"count":17},{"year":2021,"count":9},{"year":2022,"count":13},{"year":2023,"count":10},{"year":2024,"count":6},{"year":2025,"count":14},{"year":2026,"count":5}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0198714&type=printable","host_type":"journal"},{"url":"https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0198714&type=printable","host_type":"publisher"},{"url":"http://dx.plos.org/10.1371/journal.pone.0198714","host_type":"publisher"},{"url":"https://doi.org/10.1371/journal.pone.0198714","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/30222773","host_type":"repository"},{"url":"https://doaj.org/article/5ae8af2eafd846b1ad332b032c8f37da","host_type":"repository"},{"url":"http://europepmc.org/pmc/articles/PMC6141065","host_type":"repository"},{"url":"https://figshare.com/articles/dataset/Highly_multiplexed_genome_engineering_using_CRISPR_Cas9_gRNA_arrays/7095167","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/6141065","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC6141065","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC6141065?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["CRISPR and Genetic Engineering","RNA Interference and Gene Delivery","Advanced biosensing and bioanalysis techniques","CRISPR-Associated Protein 9","CRISPR-Cas Systems","Clustered Regularly Interspaced Short Palindromic Repeats","Gene Editing","HEK293 Cells","Humans","Microarray Analysis","Plasmids","Promoter Regions, Genetic","RNA, Guide, CRISPR-Cas Systems"],"mesh_terms":["Gene Editing","CRISPR-Associated Protein 9","RNA, Guide, CRISPR-Cas Systems","Humans","Plasmids","Promoter Regions, Genetic","RNA, Guide, Kinetoplastida","Microarray Analysis","HEK293 Cells","Clustered Regularly Interspaced Short Palindromic Repeats","CRISPR-Cas Systems"],"keywords":["CRISPR","Cas9","Guide RNA","Genome engineering","Trans-activating crRNA","Computational biology","Biology","RNA polymerase III","Plasmid","Genome","Genome editing","RNA","Ribonuclease","Genetics","Gene","RNA polymerase"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-26T14:27:19.000958Z","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":[]}