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Related Concept Videos

CRISPR01:59

CRISPR

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 Short...
CRISPR and crRNAs02:53

CRISPR and crRNAs

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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
CRISPR01:59

CRISPR

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 Short...

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Related Experiment Video

Updated: Jun 9, 2026

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

Functional decoupling of crRNA enables customizable CRISPR diagnostics.

Hyungbin Park1, Jiyoung Yun1, Kyuhan Lee1

  • 1Department of Chemistry, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, Republic of Korea.

Nucleic Acids Research
|March 3, 2026
PubMed
Summary

This study presents a novel CRISPR diagnostic system that decouples target programming and cleavage activity. This innovation enhances customizability for nucleic acid testing, improving diagnostic assay design.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostic Assays

Background:

  • One-pot CRISPR diagnostics offer advanced nucleic acid testing but face design limitations.
  • CRISPR RNA (crRNA) design simultaneously determines target programming and cis-cleavage activity.
  • Optimizing cleavage for isothermal amplification necessitates altering crRNA sequences, restricting target compatibility and assay flexibility.

Purpose of the Study:

  • To develop a customizable one-pot CRISPR diagnostic system by decoupling crRNA functions.
  • To enable independent control over target programming and cis-cleavage activity.
  • To overcome limitations in current CRISPR diagnostic design for flexible assay development.

Main Methods:

  • Decoupled the dual functions of crRNA design by independently controlling target programming and cis-cleavage activity.
  • Maintained target programming via the crRNA sequence.
  • Regulated cis-cleavage activity by modulating the reaction energy barrier using a crRNA-complementary RNA oligonucleotide.
  • Ensured cis-cleavage activity matched isothermal amplification conditions irrespective of the target sequence.

Main Results:

  • Achieved independent control over target programming and cis-cleavage activity in a one-pot CRISPR system.
  • Demonstrated that cis-cleavage activity can be matched to isothermal amplification without altering the crRNA sequence.
  • Validated the system's clinical applicability using 120 patient-derived samples.
  • Attained sensitivity and specificity comparable to quantitative polymerase chain reaction (qPCR).

Conclusions:

  • Resolved a fundamental constraint in CRISPR diagnostic design by decoupling crRNA functions.
  • Established a customizable and clinically deployable platform for next-generation nucleic acid testing.
  • This approach enhances the flexibility and applicability of CRISPR-based diagnostic assays.