CRISPR-Cas-amplified urinary biomarkers for multiplexed and portable cancer diagnostics
Authors:
- Liangliang HaoLHLiangliang Hao
- Renee T. ZhaoRZRenee T. Zhao
- Edward Kah Wei TanETEdward Kah Wei Tan
- Qian ZhongQZQian Zhong
- Nour Saida HarzallahNHNour Saida Harzallah
- Chayanon NgambenjawongCNChayanon Ngambenjawong
- Henry KoHKHenry Ko
- Heather E. FlemingHFHeather E. Fleming
- Sangeeta N. BhatiaSBSangeeta N. Bhatia
Top Cited Papers
Open Access
- 24 April 2023
- journal article
- research article
- Published by Springer Nature in Nature Nanotechnology
- Vol. 18 (7) , 798-807
Abstract
Synthetic biomarkers, bioengineered sensors that generate molecular reporters in diseased microenvironments, represent an emerging paradigm in precision diagnostics. Despite the utility of DNA barcodes as a multiplexing tool, their susceptibility to nucleases in vivo has limited their utility. Here we exploit chemically stabilized nucleic acids to multiplex synthetic biomarkers and produce diagnostic signals in biofluids that can be ‘read out’ via CRISPR nucleases. The strategy relies on microenvironmental endopeptidase to trigger the release of nucleic acid barcodes and polymerase-amplification-free, CRISPR-Cas-mediated barcode detection in unprocessed urine. Our data suggest that DNA-encoded nanosensors can non-invasively detect and differentiate disease states in transplanted and autochthonous murine cancer models. We also demonstrate that CRISPR-Cas amplification can be harnessed to convert the readout to a point-of-care paper diagnostic tool. Finally, we employ a microfluidic platform for densely multiplexed, CRISPR-mediated DNA barcode readout that can potentially evaluate complex human diseases rapidly and guide therapeutic decisions.Keywords
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Funding Information
- U.S. Department of Health & Human Services | NIH | National Cancer Institute (K99CA237861, P30-CA14051, P30-CA14051, P30-CA14051, P30-CA14051, P30-CA14051, P30-CA14051, P30-CA14051, P30-CA14051)
- U.S. Department of Health & Human Services | NIH | National Institute of Environmental Health Sciences (P30-ES002109, P30-ES002109, P30-ES002109, P30-ES002109, P30-ES002109, P30-ES002109, P30-ES002109, P30-ES002109)
- Howard Hughes Medical Institute
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