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Accelerated RNA detection using tandem CRISPR nucleases
Tina Y. Liu; Gavin J. Knott; Dylan C.J. Smock; John J. Desmarais; Sungmin Son; Abdul Bhuiya; Shrutee Jakhanwal; Noam Prywes; Shreeya Agrawal; María Díaz de León Derby; Neil A. Switz; Maxim Armstrong; Andrew R. Harris; Emeric J. Charles; Brittney W. Thornton; Parinaz Fozouni; Jeffrey Shu; Stephanie I. Stephens; G. Renuka Kumar; Chunyu Zhao; Amanda Mok; Anthony T. Iavarone; Arturo M. Escajeda; Roger McIntosh; Shin E. Kim; Eli J. Dugan; - IGI Testing Consortium; Katherine S. Pollard; Ming X. Tan; Melanie Ott; Daniel A. Fletcher; Liana F. Lareau; Patrick D. Hsu; David F. Savage; Jennifer A. Doudna.
Afiliação
  • Tina Y. Liu; University of California, Berkeley, Berkeley, CA, USA
  • Gavin J. Knott; Monash University, VIC 3800, Australia
  • Dylan C.J. Smock; University of California, Berkeley, Berkeley, CA, USA
  • John J. Desmarais; University of California, Berkeley, Berkeley, CA, USA
  • Sungmin Son; University of California, Berkeley, Berkeley, CA, USA
  • Abdul Bhuiya; University of California, Berkeley, Berkeley, CA, USA
  • Shrutee Jakhanwal; University of California, Berkeley, Berkeley, CA, USA
  • Noam Prywes; University of California, Berkeley, Berkeley, CA, USA
  • Shreeya Agrawal; University of California, Berkeley, Berkeley, CA, USA
  • María Díaz de León Derby; University of California, Berkeley, Berkeley, CA, USA
  • Neil A. Switz; San José State University, San José, CA, USA
  • Maxim Armstrong; Lawrence Berkeley National Laboratory, Berkeley, CA, USA
  • Andrew R. Harris; University of California, Berkeley, Berkeley, CA, USA
  • Emeric J. Charles; University of California, Berkeley, Berkeley, CA, USA
  • Brittney W. Thornton; University of California, Berkeley, Berkeley, CA, USA
  • Parinaz Fozouni; Gladstone Institutes, San Francisco, CA, USA
  • Jeffrey Shu; Gladstone Institutes, San Francisco, CA, USA
  • Stephanie I. Stephens; Gladstone Institutes, San Francisco, CA, USA
  • G. Renuka Kumar; Gladstone Institutes, San Francisco, CA, USA
  • Chunyu Zhao; Chan-Zuckerberg Biohub, San Francisco, CA, USA
  • Amanda Mok; University of California, Berkeley, Berkeley, CA, USA
  • Anthony T. Iavarone; University of California, Berkeley, Berkeley, CA, USA
  • Arturo M. Escajeda; Wainamics Inc., Pleasanton, CA, USA
  • Roger McIntosh; Wainamics Inc., Pleasanton, CA, USA
  • Shin E. Kim; University of California, Berkeley, Berkeley, CA, USA
  • Eli J. Dugan; University of California, Berkeley, Berkeley, CA, USA
  • - IGI Testing Consortium;
  • Katherine S. Pollard; Gladstone Institutes, San Francisco, CA, USA
  • Ming X. Tan; Wainamics Inc., Pleasanton, CA, USA
  • Melanie Ott; Gladstone Institutes, San Francisco, CA, USA
  • Daniel A. Fletcher; University of California, Berkeley, Berkeley, CA, USA
  • Liana F. Lareau; University of California, Berkeley, Berkeley, CA, USA
  • Patrick D. Hsu; University of California, Berkeley, Berkeley, CA, USA
  • David F. Savage; University of California, Berkeley, Berkeley, CA, USA
  • Jennifer A. Doudna; University of California, Berkeley, Berkeley, CA, USA
Preprint em En | PREPRINT-MEDRXIV | ID: ppmedrxiv-21253328
Artigo de periódico
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ABSTRACT
Direct, amplification-free detection of RNA has the potential to transform molecular diagnostics by enabling simple on-site analysis of human or environmental samples. CRISPR-Cas nucleases offer programmable RNA-guided recognition of RNA that triggers cleavage and release of a fluorescent reporter molecule1,2, but long reaction times hamper sensitivity and speed when applied to point-of-care testing. Here we show that unrelated CRISPR nucleases can be deployed in tandem to provide both direct RNA sensing and rapid signal generation, thus enabling robust detection of [~]30 RNA copies/microliter in 20 minutes. Combining RNA-guided Cas13 and Csm6 with a chemically stabilized activator creates a one-step assay that detected SARS-CoV-2 RNA from nasopharyngeal samples with PCR-derived Ct values up to 29 in microfluidic chips, using a compact imaging system. This Fast Integrated Nuclease Detection In Tandem (FIND-IT) approach enables direct RNA detection in a format amenable to point-of-care infection diagnosis, as well as to a wide range of other diagnostic or research applications.
Licença
cc_by_nc_nd
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Texto completo: 1 Coleções: 09-preprints Base de dados: PREPRINT-MEDRXIV Tipo de estudo: Diagnostic_studies Idioma: En Ano de publicação: 2021 Tipo de documento: Preprint
Texto completo
Adicionar na Minha BVS
Imprimir
XML
Buscar no Google
Texto completo: 1 Coleções: 09-preprints Base de dados: PREPRINT-MEDRXIV Tipo de estudo: Diagnostic_studies Idioma: En Ano de publicação: 2021 Tipo de documento: Preprint