{"doi":"10.1002/advs.202513439","title":"CRISPR in MOF Formulation with Enhanced Stability, Activity, and Altered PAM Specificity for Broad‐Spectrum Diagnosis of Bacterial Sepsis","abstract":"Abstract Sepsis is a life‐threatening condition caused by polymicrobial infections and remains a global health emergency that requires rapid and broad‐spectrum diagnostics. Existing CRISPR‐based assays face two major limitations that restrict their application for sepsis: narrow protospacer adjacent motif (PAM) site compatibility and poor enzyme stability under clinical and environmental stresses. A modular diagnostic platform is presented, CRISPR‐FLEXMO (CRISPR with flexible PAM in metal‐organic framework encapsulation, MOF), which integrates a PAM‐relaxed Cas12a variant (K607R) with a manganese‐coordinated MOF (Mn‐MOF) for stable and specific detection of sepsis‐causing bacteria. The system targets a conserved region upstream of the Shine‐Dalgarno sequence in the 16S rRNA gene containing a universal TTCC PAM, enabling broad‐spectrum detection with a single universal primer pair across Gram‐negative and Gram‐positive pathogens. The K607R variant shows enhanced cis ‐ and trans ‐cleavage activity, while Mn‐MOF encapsulation maintains enzyme functionality under ambient, thermal, and chaotropic stress. The assay detects as low as 10 CFU mL −1 in bacterial lysates following amplification and achieves 100% sensitivity and specificity in serum samples from 15 sepsis patients and 3 healthy individuals, with no cross‐reactivity to six respiratory viruses. The platform retains over 78% activity after 12 weeks of room‐temperature storage, offering a field‐deployable CRISPR diagnostic solution for next‐generation infectious disease detection.","journal":"Advanced Science","year":2025,"id":548883,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9537,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1442564,"name":"Zilong Liu","orcid":"0009-0009-9567-2902","position":1,"is_corresponding":false},{"id":260349,"name":"Meera G. Nair","orcid":"0000-0002-1807-5161","position":2,"is_corresponding":false},{"id":1029302,"name":"Juhong Chen","orcid":"0000-0002-6484-2739","position":3,"is_corresponding":false},{"id":1371797,"name":"Tathagata Pal","orcid":"0000-0002-2473-6974","position":0,"is_corresponding":true}],"reference_count":42,"raw_metadata":null,"created_at":"2026-07-19T02:54:03.053965Z","pmid":"41276946","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":[]}