RI-augmented TdT-CRISPR fluorescent biosensor for sensitive quantification of DNA breakage in CT-irradiated human...

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Genotoxicology

Background:

• Sperm DNA damage is a critical factor affecting male fertility.
• Assessing radiation-induced DNA damage is crucial for reproductive health.
• Current methods like the DNA Fragmentation Index (DFI) have limitations in sensitivity.

Purpose of the Study:

• To develop and validate a highly sensitive sensor for detecting radiation-induced DNA damage in sperm.
• To compare the performance of the novel sensor against conventional DFI methods.

Main Methods:

• Development of a refractive index-enhanced TdT-CRISPR sensor.
• Utilizing terminal deoxynucleotidyl transferase (TdT) and CRISPR technology for DNA damage detection.
• Quantification of DNA damage in sperm samples exposed to radiation.

Main Results:

• The RI-enhanced TdT-CRISPR sensor demonstrated a linear detection range of 0.001-0.4 nM.
• Achieved a highly sensitive limit of detection at 0.12 pM.
• The sensor precisely detected trace levels of radiation-induced DNA damage, outperforming the DFI method.

Conclusions:

• The RI-enhanced TdT-CRISPR sensor offers superior sensitivity and precision for detecting radiation-induced sperm DNA damage.
• This novel sensor represents a significant advancement over conventional methods for assessing male reproductive genotoxicity.