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. 2023 Jan 16;51(4):e24. doi: 10.1093/nar/gkac1252

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Design and evaluation of miRNA-responsive circRNA switches. (A) Evaluation by co-transfecting miRNA mimics in HEK293FT cells. Relative EGFP Expression was calculated by normalizing the sample without a miRNA mimic (magenta). 0.3 pmol of each reporter mRNA and transfection control mRNA were transfected. 0.25 pmol of specific miRNA mimic or Negative Control mimic was co-transfected for evaluation. (B) The fold-change of each miRNA-responsive circRNAs was evaluated in (A). The ON state was determined as the sample without miRNA mimic. (C) Endogenous miRNA detection by miRNA-responsive circRNAs in HEK293FT, HeLa and A549. Relative EGFP Expression was calculated by normalizing the sample with a specific miRNA inhibitor (cyan). 0.3 pmol of each reporter mRNA and transfection control mRNA were transfected. 1 pmol (hsa-miR-17-5p, HEK293FT), 2 pmol (hsa-miR-21–5p, HeLa) or 4 pmol (hsa-miR-21-5p, A549) of specific miRNA inhibitor or Negative Control inhibitor was co-transfected for evaluation. (D) The fold-change of each endogenous miRNA-responsive circRNA was evaluated in Figure 4C and fluorescent images of miRNA-responsive circRNA switches showing the best fold change. The ON state was determined as the sample with a specific miRNA inhibitor. The scale bar at the fluorescent images indicates 200 μm. Levels of significance are denoted as *P < 0.05, **P < 0.01, ***P < 0.001 (Dunnett's test). N.S. means non-significant (P >0.05). All data in this figure are presented as mean ± SD, n = 3.