Evidence map›Paper›PMID 37160120›Full record

ArticleCell reports. Medicine2023

Engineering highly thermostable Cas12b via de novo structural analyses for one-pot detection of nucleic acids.

Long T Nguyen, Santosh R Rananaware, Lilia G Yang, Nicolas C Macaluso, Julio E Ocana-Ortiz, Katelyn S Meister, Brianna L M Pizzano, Luke Samuel W Sandoval, Raymond C Hautamaki, Zoe R Fang and 9 more

Open access · goldAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Cell reports. Medicine, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers, 1 of them a synthesis that pooled it.

0numbers the graph read from it
0cells of the map it votes in
27citing papers in PubMed, 1 pooled it
9.1field-weighted citation impact, top 2% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

27 citing papers in PubMed, 1 synthesis or guideline pooled it, 59 citations in OpenAlex.

  1. Pooled it
  2. Review
  3. Review
  4. Review
  5. Article
  6. Article
  7. Review
  8. Review
  9. Review
  10. CRISPR/Cas diagnostics forFrontiers in bioengineering and biotechnology · 2026
    Review
  11. Review
  12. Simultaneous molecular detection ofFrontiers in cellular and infection microbiology · 2026
    Article
  13. Article
  14. Review
  15. Review
  16. Article
  17. Article
  18. Review
  19. Article
  20. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

19 authors at 3 institutions in 2 countries.

Long T NguyenDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Santosh R RananawareDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Lilia G YangDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Nicolas C MacalusoDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Julio E Ocana-OrtizDepartment of Chemical Engineering, University of Puerto Rico, Mayagüez, PR, USA.
Katelyn S MeisterDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Brianna L M PizzanoDepartment of Agricultural and Biological Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Luke Samuel W SandovalDepartment of Biology, College of Liberal Arts and Sciences, University of Florida, Gainesville, FL, USA.
Raymond C HautamakiDepartment of Microbiology and Cell Science, College of Agricultural and Life Sciences, University of Florida, Gainesville, FL, USA.
Zoe R FangDepartment of Microbiology and Cell Science, College of Agricultural and Life Sciences, University of Florida, Gainesville, FL, USA.
Sara M JosephDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Grace M ShoemakerDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Dylan R CarmanDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA.
Liwei ChangDepartment of Chemistry and Quantum Theory Project, College of Liberal Arts and Sciences, University of Florida, Gainesville, FL, USA.
Noah R RakestrawDepartment of Graduate Education, College of Medicine, University of Florida, Gainesville, FL, USA.
Jon F ZacharyDepartment of Graduate Education, College of Medicine, University of Florida, Gainesville, FL, USA.
Sebastian GuerraGenetics Institute, College of Medicine, University of Florida, Gainesville, FL, USA.
Alberto PerezDepartment of Chemistry and Quantum Theory Project, College of Liberal Arts and Sciences, University of Florida, Gainesville, FL, USA.
Piyush K JainDepartment of Chemical Engineering, Herbert Wertheim College of Engineering, University of Florida, Gainesville, FL, USA; Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, USA; UF Health Cancer Center, University of Florida, Gainesville, FL, USA. Electronic address: jainp@ufl.edu.
University of Florida · USUniversity of Florida Health · USUniversity of Puerto Rico-Mayaguez · PR

Funding

Discovery and engineering of CRISPR/Cas systemsR35GM147788 · NIGMS · UNIVERSITY OF FLORIDA · PI Piyush K Jain · 2022 to 2026
$1.8M
Rapid detection of Hepatitis C virus using CRISPR/CasR21AI156321 · NIAID · UNIVERSITY OF FLORIDA · PI JAIN, PIYUSH K · 2021 to 2022
$419k
Rapid point-of-care detection of Hepatitis C viral RNA using multiplexed CRISPR/Cas platformsR21AI168795 · NIAID · UNIVERSITY OF FLORIDA · PI JAIN, PIYUSH K · 2022 to 2023
$419k
NIAID NIH HHS R21 AI156321NIAID NIH HHS R21 AI168795NIGMS NIH HHS R35 GM147788
6 · The paper itself

Abstract

CRISPR-Cas-based diagnostics have the potential to elevate nucleic acid detection. CRISPR-Cas systems can be combined with a pre-amplification step in a one-pot reaction to simplify the workflow and reduce carryover contamination. Here, we report an engineered Cas12b with improved thermostability that falls within the optimal temperature range (60°C-65°C) of reverse transcription-loop-mediated isothermal amplification (RT-LAMP). Using de novo structural analyses, we introduce mutations to wild-type BrCas12b to tighten its hydrophobic cores, thereby enhancing thermostability. The one-pot detection assay utilizing the engineered BrCas12b, called SPLENDID (single-pot LAMP-mediated engineered BrCas12b for nucleic acid detection of infectious diseases), exhibits robust trans-cleavage activity up to 67°C in a one-pot setting. We validate SPLENDID clinically in 80 serum samples for hepatitis C virus (HCV) and 66 saliva samples for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with high specificity and accuracy. We obtain results in as little as 20 min, and with the extraction process, the entire assay can be performed within an hour.

Indexed as

COVID-19Nucleic AcidsCOVID-19 TestingCRISPR-Cas SystemsHumansSARS-CoV-2Nucleic AcidsCas12bCOVID-19CRISPRdiagnosticshepatitis Cone-pot detectionpoint-of-care diagnosticsprotein engineeringRT-LAMPthermal stability

Identifiers

PMID37160120
PMCPMC10213852
OpenAlexW4376121457

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.