Split aptamer regulated CRISPR/Cas12a biosensor for 17β-estradiol through a gap-enhanced Raman tags based lateral flow...

Area of Science:

• Biotechnology
• Biosensor Development
• Molecular Diagnostics

Background:

• CRISPR/Cas systems offer potential for non-nucleic acid detection.
• Developing sensitive and user-friendly biosensors for small molecules like 17β-estradiol is crucial.

Purpose of the Study:

• To develop a split aptamer-regulated CRISPR/Cas12a biosensor for 17β-estradiol detection.
• To integrate CRISPR/Cas12a with gap-enhanced Raman tags (GERT) for a lateral flow assay.
• To achieve visible and ultra-sensitive detection of 17β-estradiol.

Main Methods:

• Utilized a split aptamer strategy to regulate CRISPR/Cas12a activity based on 17β-estradiol binding.
• Integrated CRISPR/Cas12a trans-cleavage with GERT-based lateral flow assay (LFA).
• Developed a dual-mode (visible and SERS) detection system.

Main Results:

• Achieved visible recognition of 17β-estradiol down to 10 pM.
• Quantified 17β-estradiol with a detection limit of 180 fM using SERS.
• Demonstrated a detection sensitivity at least 1000-fold higher than previous immunoassay LFA strategies.

Conclusions:

• The developed split aptamer-regulated CRISPR/Cas12a-GERT LFA provides a novel approach for non-nucleic acid detection.
• This assay enables ultra-sensitive, visible, and user-friendly detection of small-molecule targets.
• Offers a promising platform for developing advanced biosensors for various applications.