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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Related Experiment Video

Updated: Jul 17, 2025

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
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Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

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Programmable RNA detection with CRISPR-Cas12a.

Santosh R Rananaware1, Emma K Vesco1,2, Grace M Shoemaker1

  • 1Department of Chemical Engineering, University of Florida, Gainesville, FL, USA.

Nature Communications
|September 5, 2023
PubMed
Summary
This summary is machine-generated.

Split Activator for Highly Accessible RNA Analysis (SAHARA) uses Cas12a for sensitive RNA detection without amplification. This CRISPR-Cas12a based platform offers high specificity and multiplexing capabilities for nucleic acid analysis.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • CRISPR-Cas12a is a DNA-cleaving enzyme used in diagnostics.
  • Cas12a's trans-cleavage activity can be activated by split activators.
  • Current RNA detection methods often require amplification or reverse-transcription.

Purpose of the Study:

  • To develop a novel CRISPR-Cas12a based method for direct RNA detection.
  • To enhance the specificity and multiplexing capabilities of Cas12a for nucleic acid analysis.
  • To create a sensitive diagnostic tool for RNA detection without complex sample preparation.

Main Methods:

  • Utilized split activators to enable Cas12a trans-cleavage on RNA targets.
  • Developed the Split Activator for Highly Accessible RNA Analysis (SAHARA) method.
  • Designed crRNAs with specific PAM-proximal seed regions for DNA targeting.

Main Results:

  • SAHARA detects picomolar concentrations of RNA without amplification or reverse-transcription.
  • The method demonstrates enhanced specificity towards point mutations compared to wild-type Cas12a.
  • SAHARA enables multiplexed detection of multiple RNA and DNA targets using pooled crRNA/Cas12a arrays.

Conclusions:

  • SAHARA is a simple and powerful nucleic acid detection platform based on Cas12a.
  • The platform offers high sensitivity, specificity, and multiplexing potential for RNA and DNA analysis.
  • SAHARA can be expanded to other CRISPR-Cas enzymes and applied in various diagnostic settings.