{"doi":"10.3791/66189","title":"CRISPR-Cas-mediated Multianalyte Synthetic Urine Biomarker Test for Portable Diagnostics","abstract":null,"journal":"Journal of Visualized Experiments","year":2023,"id":608324,"datarank":0.20794415416798362,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"self_citation_contribution":0.20794415416798362,"citation_network_contribution":0.0,"self_endowment_contribution":0.20794415416798362,"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":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":763432,"name":"Feiyang Deng","orcid":"0000-0003-0598-4259","position":1,"is_corresponding":false},{"id":247584,"name":"Renee T. 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Synthetic biomarkers generally make use of reporters that provide readable signals in the biofluid to reflect the biochemical alterations in the local disease microenvironment during disease incidence and progression. The pharmacokinetic concentration of the reporters and biochemical amplification of the disease signal are paramount to achieving high sensitivity and specificity in a diagnostic test. Here, a cancer diagnostic platform is built using one format of synthetic biomarkers: activity-based nanosensors carrying chemically stabilized DNA reporters that can be liberated by aberrant proteolytic signatures in the tumor microenvironment. Synthetic DNA as a disease reporter affords multiplexing capability through its use as a barcode, allowing for the readout of multiple proteolytic signatures at once. DNA reporters released into the urine are detected using CRISPR nucleases via hybridization with CRISPR RNAs, which in turn produce a fluorescent or colorimetric signal upon enzyme activation. In this protocol, DNA-barcoded, activity-based nanosensors are constructed and their application is exemplified in a preclinical mouse model of metastatic colorectal cancer. This system is highly modifiable according to disease biology and generates multiple disease signals simultaneously, affording a comprehensive understanding of the disease characteristics through a minimally invasive process requiring only nanosensor administration, urine collection, and a paper test which enables point-of-care diagnostics.","is_dataset_classified":null,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"38145378","pmcid":"PMC10840402","openalex_id":"https://openalex.org/W4389498108","authors":[],"funders":[{"funder_name":"NCI NIH HHS","grant_id":"R00 CA237861","title":null},{"funder_name":"NIBIB NIH HHS","grant_id":"T32 EB006359","title":null},{"funder_name":"NCI NIH HHS","grant_id":"P30 CA014051","title":null},{"funder_name":"NIEHS NIH HHS","grant_id":"P30 ES002109","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"T32 GM130546","title":null},{"funder_name":"National Institutes of Health","grant_id":"5P30CA014051-17","title":"CELL BIOLOGY--DEVELOPMENTAL PROGRAM"},{"funder_name":"National Institutes of Health","grant_id":"2P30ES002109-11","title":"TOXICOLOGY CORE UNIT"},{"funder_name":"National Institutes of Health","grant_id":"5T32GM130546-04","title":"Synthetic Biology and Biotechnology (SB2) Predoctoral Training Program"},{"funder_name":"Howard Hughes Medical Institute","grant_id":"","title":null}],"total_grants":9,"fwci":0.2793,"citation_percentile":0.52005993,"influential_citations":0,"citation_trend":[{"year":2025,"count":1},{"year":2026,"count":2}],"oa_status":"green","license":null,"oa_locations":[{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC10840402/pdf/nihms-1958300.pdf","host_type":"repository"},{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC10840402/pdf/nihms-1958300.pdf","host_type":"repository"},{"url":"https://www.jove.com/t/66189/crispr-cas-mediated-multianalyte-synthetic-urine-biomarker-test-for","host_type":"publisher"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/10840402","host_type":"repository"},{"url":"https://doi.org/10.3791/66189","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/38145378","host_type":"repository"},{"url":"http://doi.org/10.3791/66189","host_type":"repository"},{"url":"https://doi.org/https://doi.org/10.3791/66189","host_type":""}],"fields_of_study":["CRISPR and Genetic Engineering","Advanced biosensing and bioanalysis techniques","Innovative Microfluidic and Catalytic Techniques Innovation","0303 health sciences","03 medical and health sciences"],"mesh_terms":["Animals","Body Fluids","DNA","Biomarkers","Urinalysis","Mice","CRISPR-Cas Systems"],"keywords":["CRISPR","Nanosensor","Computational biology","Biomarker","Multiplex","Biology","Bioinformatics","Nanotechnology","Genetics","Gene","Materials science","610","DNA","Urinalysis","Body Fluids","Mice","Cognitive sciences","Biochemistry and cell biology","Psychology","Animals","CRISPR-Cas Systems","Biomarkers"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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