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A novel biosensor for highly sensitive DNA damage detection using TdT and CRISPR-Cas12a.

Juan Wang1, Fan Zhang2, Ziyang Liu1

  • 1Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Institute of Medical Sciences, General Hospital of Ningxia Medical University, Yinchuan, 750004, China. d202081630@alumni.hust.edu.cn.

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This study introduces a sensitive fluorescence biosensor for detecting sperm DNA damage. The novel tool utilizes TdT enzyme and CRISPR-Cas12a, offering a simple and versatile method for reproductive health diagnostics.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genetics

Background:

  • Sperm DNA damage impacts male fertility and reproductive outcomes.
  • Accurate and sensitive detection methods are crucial for clinical diagnostics and reproductive medicine.

Purpose of the Study:

  • To develop a highly sensitive fluorescence biosensor for precise detection of sperm DNA damage.
  • To integrate terminal deoxynucleotidyl transferase (TdT) enzyme with CRISPR-Cas12a technology for enhanced detection capabilities.

Main Methods:

  • Development of a fluorescence biosensor system.
  • Integration of TdT enzyme for labeling DNA strand breaks.
  • Utilizing CRISPR-Cas12a collateral activity for signal amplification and detection.
  • Characterization of biosensor sensitivity and linear detection range.

Main Results:

  • Achieved a highly sensitive detection limit of 0.99 pM for sperm DNA damage.
  • Established a linear detection range from 0.001 nM to 0.2 nM.
  • Demonstrated exceptional sensitivity, simplicity, and versatility of the developed biosensor.

Conclusions:

  • The TdT and CRISPR-Cas12a integrated biosensor is a transformative tool for sperm DNA damage detection.
  • This method offers significant potential for advancing reproductive medicine and clinical diagnostics.
  • The biosensor's high sensitivity and ease of use pave the way for improved fertility assessments.