{"doi":"10.1101/2023.09.16.558090","title":"Multiplexed CRISPR gene editing in primary human islet cells with Cas9 ribonucleoprotein","abstract":"Summary Successful genome editing in primary human islets could reveal features of the genetic regulatory landscape underlying β cell function and diabetes risk. Here, we describe a CRISPR-based strategy to interrogate functions of predicted regulatory DNA elements using electroporation of a complex of Cas9 ribonucleoprotein (Cas9 RNP) and guide RNAs into primary human islet cells. We successfully targeted coding regions including the PDX1 exon 1, and non-coding DNA linked to diabetes susceptibility. CRISPR/Cas9 RNP approaches revealed genetic targets of regulation by DNA elements containing candidate diabetes risk SNPs, including an in vivo enhancer of the MPHOSPH9 gene. CRISPR/Cas9 RNP multiplexed targeting of two cis -regulatory elements linked to diabetes risk in PCSK1 , which encodes an endoprotease crucial for insulin processing, also demonstrated efficient simultaneous editing of PCSK1 regulatory elements, resulting in impaired β cell PCSK1 regulation and insulin secretion. Multiplex CRISPR/Cas9 RNP provides powerful approaches to investigate and elucidate human islet cell gene regulation in health and diabetes.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":395088,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.949,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":661976,"name":"Weichen Zhao","orcid":"0000-0003-4838-7016","position":1,"is_corresponding":false},{"id":675070,"name":"Emilio Merheb","orcid":"0000-0002-1415-1075","position":2,"is_corresponding":false},{"id":1169901,"name":"Seung‐Hyun Kim","orcid":"0000-0003-1159-4521","position":3,"is_corresponding":false},{"id":30775,"name":"Alexander Marson","orcid":"0000-0002-2734-5776","position":5,"is_corresponding":false},{"id":7383,"name":"Anna L. Gloyn","orcid":"0000-0003-1205-1844","position":6,"is_corresponding":false},{"id":44518,"name":"Seung K. Kim","orcid":"0000-0001-6135-7810","position":7,"is_corresponding":false},{"id":467426,"name":"Romina J. Bevacqua","orcid":"0000-0002-3903-6508","position":0,"is_corresponding":true}],"reference_count":35,"raw_metadata":null,"created_at":"2026-07-19T01:19:18.686170Z","pmid":"37745551","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":[]}