Journal
BIOSENSORS & BIOELECTRONICS
Volume 192, Issue -, Pages -Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2021.113507
Keywords
RNA imaging; Rolling circle amplification; Photothermal nanomaterial; In situ mRNA detection; Frozen tissue section; Formalin-fixed paraffin-embedded tissues section
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Funding
- National Major Science and Tech-nology Projects [2018ZX10101001]
- PUMC Scholar fund from the Chinese Academy of Medical Sciences
- CAMS Innovation Fund for Medical Sciences [2018-I2M-1-001]
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By introducing Au-TPP and 'C' form target-specific oligonucleotide linker probes, a specific and photothermal mediated hyperbranched rolling circle amplification was successfully developed for direct in situ mRNA detection, suitable for both FFPE tissue section and frozen tissue section.
Rolling circle amplification (RCA) had the prospect of assisting clinic diagnosis with advantage in in situ mRNA detection at single cell level. However, for direct mRNA detection, RCA had relatively low detection specificity and efficiency. Here, we introduced 4-(10, 15, 20-Triphenylporphyrin-5-yl)phenylamine (TPP) modified Au nanoparticle (Au-TPP) to improve the specificity of in-situ RCA. Through photothermal effect, Au-TPP acted as the specific heat source upon irradiation of 635 nm laser. The photothermal mediated RCA would be initiated only when the Au-TPP as well as the padlock anchored adjacently on the same target mRNA. Furthermore, we introduced 'C' form target-specific oligonucleotide linker probes to make generic padlock and Au-TPP for different mRNA targets, so that for a new mRNA target one does not have to redesign the padlock and the Au-TPP probe. By these strategies, we successfully developed a specific and photothermal mediated hyperbranched rolling circle amplification for direct in situ mRNA detection, suitable for both formalin-fixed paraffin-embedded (FFPE) tissue section and frozen tissue section.
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