Split aptamer regulated CRISPR/Cas12a biosensor for 17β-estradiol through a gap-enhanced Raman tags based lateral flow strategy
- Biosens Bioelectron. 2022 Nov 1:215:114548. doi: 10.1016/j.bios.2022.114548.
- 1. Capital Medical University, Department of Toxicology, No. 10 Xitoutiao, You An Men, Beijing, 100069, PR China.
- 2. Capital Medical University, Department of Toxicology, No. 10 Xitoutiao, You An Men, Beijing, 100069, PR China. Electronic address: [email protected].
- 3. Beijing Institute of Microbiology and Epidemiology, 27 Taiping Road, 100850, Beijing, PR China. Electronic address: [email protected].
- 4. Capital Medical University, Department of Toxicology, No. 10 Xitoutiao, You An Men, Beijing, 100069, PR China. Electronic address: [email protected].
It is significant to exploit the full potential of CRISPR/Cas based biosensor for non-nucleic-acid targets. Here, we developed a split aptamer regulated CRISPR/Cas12a and gap-enhanced Raman tags based lateral flow biosensor for small-molecule target, 17β-estradiol. In this assay, one split aptamer of 17β-estradiol was designed to complement with crRNA of Cas12a so that the trans-cleavage ability of CRISPR/Cas12a can be regulated by the competitive binding of 17β-estradiol and split Aptamers. Through integration of the signal amplification ability of CRISPR/Cas12a and the ultra-sensitive gap-enhanced Raman tags based lateral flow assay, a visible-SERS dual mode determination of 17β-estradiol can be established. 17β-estradiol can be visibly recognized as low as 10 pM and accurately quantified with a detection limit of 180 fM by SERS signals, which is at least 103-fold lower than that of the previous Immunoassay lateral flow strategies. Our assay provides a novel perspective to develop split aptamer regulated CRISPR/Cas12a coupling with SERS lateral flow strips for ultrasensitive and easy-to-use non-nucleic-acid targets detection.
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Research Areas: Cancer