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

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.
Qing Li 1,  Xiaobo Li 1,  Pengyou Zhou 1,  Rui Chen 2,  Rui Xiao 3,  Yuanfeng Pang 4
Affiliations

Affiliations

  • 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].
PMID: 35870335 DOI: 10.1016/j.bios.2022.114548
Abstract

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.

Keywords

17β-estradiol; CRISPR/Cas12a; Gap-enhanced Raman tags; Lateral flow assay; Split aptamers.

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