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Related Experiment Video

Updated: Jun 29, 2025

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CRISPR-HOLMES-based NAD+ detection.

Songkuan Zhuang1,2, Tianshuai Hu2, Hongzhong Zhou2

  • 1Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Shenzhen University Medical School, Shenzhen, China.

Frontiers in Bioengineering and Biotechnology
|April 9, 2024
PubMed
Summary
This summary is machine-generated.

A new CRISPR-Cas12a based system, HOLMES(NAD+), offers rapid and convenient detection of intracellular nicotinamide adenine dinucleotide (NAD+). This method overcomes limitations of traditional techniques, enabling sensitive and specific NAD+ quantitation.

Keywords:
CRISPRCas12aHOLMESNAD+acetylation

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Intracellular nicotinamide adenine dinucleotide (NAD+) levels are crucial biomarkers for numerous diseases.
  • Conventional NAD+ detection methods are often laborious, time-consuming, and require expensive equipment.

Purpose of the Study:

  • To develop a rapid, convenient, and sensitive method for detecting intracellular NAD+.
  • To leverage CRISPR-Cas12a technology for non-nucleic acid detection applications.

Main Methods:

  • Development of a one-step CRISPR-Cas12a-based HOLMES(NAD+) system.
  • Utilized acetylated Cas12a, inactivated by AcrVA5, and reactivated by CobB using NAD+ as a cofactor.
  • Employed ssDNA reporters for fluorescence signal generation upon NAD+-dependent Cas12a reactivation.

Main Results:

  • The HOLMES(NAD+) system demonstrated high sensitivity and specificity for NAD+ detection.
  • Successfully enabled quantitative determination of intracellular NAD+ concentrations.
  • Validated the expansion of HOLMES applications beyond nucleic acid detection.

Conclusions:

  • HOLMES(NAD+) provides an efficient and rapid approach for NAD+ quantitation.
  • This system offers a valuable tool for disease research and diagnostics related to NAD+ metabolism.
  • The study expands the utility of CRISPR-Cas12a systems for detecting small molecules.