Field-deployable viral diagnostics using CRISPR-Cas13
Cameron Myhrvold,Cameron Myhrvold,Catherine A. Freije,Catherine A. Freije,Jonathan S. Gootenberg,Omar O. Abudayyeh,Hayden C. Metsky,Ann Durbin,Ann Durbin,Max J. Kellner,Amanda L Tan,Lauren M. Paul,Leda Parham,Kimberly García,Kayla G. Barnes,Kayla G. Barnes,Bridget Chak,Bridget Chak,Adriano Mondini,Maurício Lacerda Nogueira,Sharon Isern,Scott F. Michael,Ivette Lorenzana,Nathan L. Yozwiak,Nathan L. Yozwiak,Bronwyn MacInnis,Bronwyn MacInnis,Irene Bosch,Irene Bosch,Lee Gehrke,Lee Gehrke,Feng Zhang,Pardis C. Sabeti +32 more
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TLDR
The Cas13-based SHERLOCK platform can detect Zika virus and dengue virus in patient samples at concentrations as low as 1 copy per microliter and can distinguish the four DENV serotypes, as well as region-specific strains of ZIKV from the 2015–2016 pandemic.Abstract:
Mitigating global infectious disease requires diagnostic tools that are sensitive, specific, and rapidly field deployable. In this study, we demonstrate that the Cas13-based SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform can detect Zika virus (ZIKV) and dengue virus (DENV) in patient samples at concentrations as low as 1 copy per microliter. We developed HUDSON (heating unextracted diagnostic samples to obliterate nucleases), a protocol that pairs with SHERLOCK for viral detection directly from bodily fluids, enabling instrument-free DENV detection directly from patient samples inread more
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Journal ArticleDOI
CRISPR-Cas12-based detection of SARS-CoV-2.
James Paul Broughton,Xianding Deng,Guixia Yu,Clare L Fasching,Venice Servellita,Jasmeet Singh,Xin Miao,Jessica Streithorst,Andrea Granados,Alicia Sotomayor-Gonzalez,Kelsey C. Zorn,Allan Gopez,Elaine Hsu,Wei Gu,Steve Miller,Chao Yang Pan,Hugo Guevara,Debra A. Wadford,Janice S. Chen,Charles Y. Chiu +19 more
TL;DR: The CRISPR-based DETECTR assay provides a visual and faster alternative to the US Centers for Disease Control and Prevention SARS-CoV-2 real-time RT–PCR assay, with 95% positive predictive agreement and 100% negative predictive agreement.
Journal ArticleDOI
Virology, Epidemiology, Pathogenesis, and Control of COVID-19.
Yuefei Jin,Haiyan Yang,Wangquan Ji,Weidong Wu,Shuaiyin Chen,Weiguo Zhang,Weiguo Zhang,Guangcai Duan +7 more
TL;DR: The present understanding of COVID-19 is detailed and the current state of development of measures are introduced in this review to provide a comprehensive summary to public health authorities and potential readers worldwide.
Journal ArticleDOI
CRISPR-Cas guides the future of genetic engineering.
TL;DR: The basic mechanisms that set the CRISPR-Cas toolkit apart from other programmable gene-editing technologies are described, highlighting the diverse and naturally evolved systems now functionalized as biotechnologies.
Journal ArticleDOI
SHERLOCK: nucleic acid detection with CRISPR nucleases.
Max J. Kellner,Jeremy G. Koob,Jonathan S. Gootenberg,Jonathan S. Gootenberg,Jonathan S. Gootenberg,Omar O. Abudayyeh,Omar O. Abudayyeh,Omar O. Abudayyeh,Feng Zhang +8 more
TL;DR: Step-by-step instructions for setting up SHERLOCK assays with recombinase-mediated polymerase pre-amplification of DNA or RNA and subsequent Cas13- or Cas12-mediated detection via fluorescence and colorimetric readouts that provide results in <1 h with a setup time of less than 15 min are provided.
Journal ArticleDOI
CRISPR/Cas Systems towards Next-Generation Biosensing.
TL;DR: A detailed classification of CRISPR/Cas biosensing systems is provided and they have the potential to become promising candidates for next-generation diagnostic biosensing platforms.
References
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Journal ArticleDOI
Nucleic acid detection with CRISPR-Cas13a/C2c2
Jonathan S. Gootenberg,Omar O. Abudayyeh,Jeong Wook Lee,Patrick Essletzbichler,Aaron J. Dy,Aaron J. Dy,Julia Joung,Vanessa Verdine,Nina M. Donghia,Nichole M. Daringer,Catherine A. Freije,Catherine A. Freije,Cameron Myhrvold,Cameron Myhrvold,Roby P. Bhattacharyya,Jonathan Livny,Aviv Regev,Aviv Regev,Eugene V. Koonin,Deborah T. Hung,Pardis C. Sabeti,James J. Collins,Feng Zhang +22 more
TL;DR: A Cas13a-based molecular detection platform, termed Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK), is used to detect specific strains of Zika and Dengue virus, distinguish pathogenic bacteria, genotype human DNA, and identify mutations in cell-free tumor DNA.
Journal ArticleDOI
DNA Detection Using Recombination Proteins
TL;DR: RPA couples isothermal recombinase polymerase-driven primer targeting of template material with strand-displacement DNA synthesis and achieves exponential amplification with no need for pretreatment of sample DNA, thereby establishing an instrument-free DNA testing system.
Journal ArticleDOI
C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector
Omar O. Abudayyeh,Jonathan S. Gootenberg,Silvana Konermann,Silvana Konermann,Silvana Konermann,Julia Joung,Julia Joung,Julia Joung,Ian Slaymaker,Ian Slaymaker,Ian Slaymaker,David Benjamin Turitz Cox,Sergey Shmakov,Sergey Shmakov,Kira S. Makarova,Ekaterina Semenova,Leonid Minakhin,Konstantin Severinov,Konstantin Severinov,Konstantin Severinov,Aviv Regev,Aviv Regev,Eric S. Lander,Eric S. Lander,Eric S. Lander,Eugene V. Koonin,Feng Zhang +26 more
TL;DR: LshC2c2 is a RNA-guided RNase which requires the activity of its two HEPN domains, suggesting previously unidentified mechanisms of RNA targeting and degradation by CRISPR systems.
Journal ArticleDOI
Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6
Jonathan S. Gootenberg,Omar O. Abudayyeh,Max J. Kellner,Julia Joung,James J. Collins,Feng Zhang +5 more
TL;DR: ShERLOCK as discussed by the authors is a platform that combines isothermal preamplification with Cas13 to detect single molecules of RNA or DNA, which can detect Dengue or Zika virus single-stranded RNA and mutations in patient liquid biopsy samples via lateral flow.
Journal ArticleDOI
Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components
Keith Pardee,Alexander A. Green,Melissa K. Takahashi,Dana Braff,Dana Braff,Dana Braff,Guillaume Lambert,Guillaume Lambert,Jeong Wook Lee,Tom Ferrante,Duo Ma,Nina M. Donghia,Melina Fan,Nichole M. Daringer,Irene Bosch,Dawn M. Dudley,David H. O’Connor,Lee Gehrke,Lee Gehrke,James J. Collins +19 more
TL;DR: A pipeline for the rapid design, assembly, and validation of cell-free, paper-based sensors for the detection of the Zika virus RNA genome is reported, which detect clinically relevant concentrations of Zika virus sequences and demonstrate specificity against closely related Dengue virus sequences.
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