CRISPR-enabled point-of-care genotyping for APOL1 genetic risk assessment.

Autor:	Greensmith R; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.; Department of Nephrology and Medical Intensive Care, Charité Universitätsmedizin Berlin, Berlin, Germany., Lape IT; Center for Transplantation Sciences, Massachusetts General Hospital/Harvard Medical School, Boston, MA, USA., Riella CV; Nephrology Division, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA, 02215, USA.; Harvard Medical School, Boston, MA, 02115, USA., Schubert AJ; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.; Department of Nephrology and Medical Intensive Care, Charité Universitätsmedizin Berlin, Berlin, Germany.; Berlin Institute of Health, Berlin, Germany., Metzger JJ; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany., Dighe AS; Harvard Medical School, Boston, MA, 02115, USA.; Department of Pathology, Massachusetts General Hospital, Boston, MA, USA., Tan X; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Division of Gastroenterology, Massachusetts General Hospital, Boston, MA, USA.; Institute for Medical Engineering and Science and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.; Infectious Disease and Microbiome Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA., Hemmer B; Department of Neurology, Klinikum rechts der Isar, Technical University of Munich, Munich, Germany.; Munich Cluster for Systems Neurology (SyNergy), Munich, Germany., Rau J; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany., Wendlinger S; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.; Department of Nephrology and Medical Intensive Care, Charité Universitätsmedizin Berlin, Berlin, Germany., Diederich N; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.; Department of Nephrology and Medical Intensive Care, Charité Universitätsmedizin Berlin, Berlin, Germany., Schütz A; Protein Production & Characterization, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 13125, Berlin, Germany., Riella LV; Center for Transplantation Sciences, Massachusetts General Hospital/Harvard Medical School, Boston, MA, USA. lriella@mgh.harvard.edu.; Department of Surgery, Massachusetts General Hospital/Harvard Medical School, Boston, MA, USA. lriella@mgh.harvard.edu.; Division of Nephrology, Massachusetts General Hospital/Harvard Medical School, Boston, MA, USA. lriella@mgh.harvard.edu., Kaminski MM; Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany. michael.kaminski@mdc-berlin.de.; Department of Nephrology and Medical Intensive Care, Charité Universitätsmedizin Berlin, Berlin, Germany. michael.kaminski@mdc-berlin.de.; Berlin Institute of Health, Berlin, Germany. michael.kaminski@mdc-berlin.de.
Jazyk:	angličtina
Zdroj:	EMBO molecular medicine [EMBO Mol Med] 2024 Oct; Vol. 16 (10), pp. 2619-2637. Date of Electronic Publication: 2024 Sep 13.
DOI:	10.1038/s44321-024-00126-x
Abstrakt:	Detecting genetic variants enables risk factor identification, disease screening, and initiation of preventative therapeutics. However, current methods, relying on hybridization or sequencing, are unsuitable for point-of-care settings. In contrast, CRISPR-based-diagnostics offer high sensitivity and specificity for point-of-care applications. While these methods have predominantly been used for pathogen sensing, their utilization for genotyping is limited. Here, we report a multiplexed CRISPR-based genotyping assay using LwaCas13a, PsmCas13b, and LbaCas12a, enabling the simultaneous detection of six genotypes. We applied this assay to identify genetic variants in the APOL1 gene prevalent among African Americans, which are associated with an 8-30-fold increase in the risk of developing kidney disease. Machine learning facilitated robust analysis across a multicenter clinical cohort of more than 100 patients, accurately identifying their genotypes. In addition, we optimized the readout using a multi-analyte lateral-flow assay demonstrating the ability for simplified genotype determination of clinical samples. Our CRISPR-based genotyping assay enables cost-effective point-of-care genetic variant detection due to its simplicity, versatility, and fast readout. (© 2024. The Author(s).)
Databáze:	MEDLINE
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