2. Academic Validation
  3. Bimetallic-satellite enhanced SERS reporter integrated with CRISPR-Cas12a for ultrasensitive biological rhythm monitoring

Bimetallic-satellite enhanced SERS reporter integrated with CRISPR-Cas12a for ultrasensitive biological rhythm monitoring

  • Anal Chim Acta. 2025 Nov 8:1374:344544. doi: 10.1016/j.aca.2025.344544.
Hong Zhang 1,  Xuekun Bai 2,  Chenxi Wang 2,  Wei Pang 2,  Jiaxin Zhang 2,  Junhuai Song 3,  Huanying Zhou 4,  Zhixian Gao 5,  Bin Zheng 6
Affiliations

Affiliations

  • 1 Department of Biochemistry and Molecular Biology, Key Laboratory of Neural and Vascular Biology, Ministry of Education, Hebei Key Laboratory of Cardiovascular Homeostasis and Aging, Hebei Medical University, Shijiazhuang, 050017, China; Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Military Medical Sciences Academy, Academy of Military Sciences, Tianjin, 300050, China.
  • 2 Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Military Medical Sciences Academy, Academy of Military Sciences, Tianjin, 300050, China.
  • 3 Department of Biochemistry and Molecular Biology, Key Laboratory of Neural and Vascular Biology, Ministry of Education, Hebei Key Laboratory of Cardiovascular Homeostasis and Aging, Hebei Medical University, Shijiazhuang, 050017, China.
  • 4 Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Military Medical Sciences Academy, Academy of Military Sciences, Tianjin, 300050, China. Electronic address: [email protected].
  • 5 Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Military Medical Sciences Academy, Academy of Military Sciences, Tianjin, 300050, China. Electronic address: [email protected].
  • 6 Department of Biochemistry and Molecular Biology, Key Laboratory of Neural and Vascular Biology, Ministry of Education, Hebei Key Laboratory of Cardiovascular Homeostasis and Aging, Hebei Medical University, Shijiazhuang, 050017, China. Electronic address: [email protected].
PMID: 40983435 DOI: 10.1016/j.aca.2025.344544
Abstract

Background: Biorhythmic played an important role in physical health. Biorhythmic disorders led to various health issue, including cardiovascular diseases and metabolic disorders. Early diagnosis of biorhythmic disorders was crucial in slowing the progression of disease. Therefore, it was important to develop technologies for detecting biorhythmic and their disorder markers. This study selected HMGB1(High mobility group box 1), BMAL1 (Brain and Muscle Aryl Hydrocarbon Receptor nuclear translocator-like 1), and MICU (mitochondrial calcium uptake 1) mRNA for detection.

Results: CRISPR/Cas12a/SERS (surface-enhanced Raman spectroscopy) ultrasensitive biosensing platform was developed for the detection of biorhythmic markers. The biosensor was partly an ultrabright Raman signal reporter (on) of a novel satellite structure formed by gold/silver bimetallic nanoparticles. This novel satellite structure utilizes the potential difference between gold and silver nanoparticles to promote the transfer of electrons from the core structure to the AuNPs and MBA (4-mercaptobenzoic acid) molecules, while enhancing the electromagnetic field distribution. Compared with the Raman signal of the satellite structure composed of Au/Au bimetallic, the Raman signal of the satellite structure in this study is increased by about 10 times. CRISPR/Cas12a recognized the target molecule and activated trans-cleavage activity, cleaving the DNA/RNA hairpin structure and releasing RNA2∗. RNA2∗ caused the satellite structure to disperse, significantly attenuating the ultra-strong Raman signal (off), thereby enabling quantitative detection of the target nucleic acid. The LOD for three targets were of less than 1 fM.

Significance: In summary, the CRISPR/Cas12a/SERS biosensing platform enables mRNA detection without preamplification. This biosensing platform can detect target molecules at the fM level, achieving ultra-sensitive detection. In addition, the satellite structure exhibits long-term stability, providing a theoretical basis for the practical application of this biosensing platform.

Keywords

Bimetallic satellite structure; Biorhythms; CRISPR/Cas12a; Nucleic acid detection; SERS.

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