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Thermo Fisher mirvana mirna mimic negative control #1, scrambled
Target genes of miR-124-3p, miR-23b-3p and shared by both <t>miRNAs.</t> (A) The Venn diagram shows the target mRNAs of miR-124-3p and miR-23b-3p recorded in miRDB and TargetScan. (B) The table shows the name of the 136 genes predicted as targets shared by miR-124-3p and miR-23b-3p. miR, microRNA; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Target genes of miR-124-3p, miR-23b-3p and shared by both <t>miRNAs.</t> (A) The Venn diagram shows the target mRNAs of miR-124-3p and miR-23b-3p recorded in miRDB and TargetScan. (B) The table shows the name of the 136 genes predicted as targets shared by miR-124-3p and miR-23b-3p. miR, microRNA; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Mirvana Mirna Mimic Negative Control #1, Scrambled, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mirna+mimic+negative+control/mirvana+mirna+mimic+negative+control++1++scrambled/pmc12218049-84-4-11
Average 90 stars, based on 1 article reviews

mirvana mirna mimic negative control #1, scrambled - by Bioz Stars, 2026-09

90/100 stars

Images

1) Product Images from "SLC7A1 , SGK1 and HMGB2 are overexpressed in cervical cancer tissues and the miR-23b-3p/HMGB2 axis regulates cell migration and invasion"

Article Title: SLC7A1 , SGK1 and HMGB2 are overexpressed in cervical cancer tissues and the miR-23b-3p/HMGB2 axis regulates cell migration and invasion

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2025.13600

Target genes of miR-124-3p, miR-23b-3p and shared by both miRNAs. (A) The Venn diagram shows the target mRNAs of miR-124-3p and miR-23b-3p recorded in miRDB and TargetScan. (B) The table shows the name of the 136 genes predicted as targets shared by miR-124-3p and miR-23b-3p. miR, microRNA; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Figure Legend Snippet: Target genes of miR-124-3p, miR-23b-3p and shared by both miRNAs. (A) The Venn diagram shows the target mRNAs of miR-124-3p and miR-23b-3p recorded in miRDB and TargetScan. (B) The table shows the name of the 136 genes predicted as targets shared by miR-124-3p and miR-23b-3p. miR, microRNA; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Techniques Used:

other:

Article Title: Transmission of reduced levels of miR-34/449 from sperm to preimplantation embryos is a key step in the transgenerational epigenetic inheritance of the effects of paternal chronic social instability stress.
Article Snippet: Each miRNA (mimic miR-34c, 50pmole, #MC11039, or mimic negative control, #4464058, Thermofisher) was added to 5 l R buffer (Invitrogen) and incubated for 10 minutes prior to transfection.

Article Title: Endothelial extracellular vesicles enhance vascular self-assembly in engineered human cardiac tissues.
Article Snippet: Accepted Manuscript is “the version of the article accepted for publication including all changes made as a result of the peer review process, and which may also include the addition to the article by IOP Publishing of a header, an article ID, a cover sheet and/or an ‘Accepted Manuscript’ watermark, but excluding any other editing, typesetting or other changes made by IOP Publishing and/or its licensors” This Accepted Manuscript is © 2024 The Author(s).. Published by IOP Publishing Ltd.

Article Title: Immune escape of multiple myeloma cells results from low miR29b and the ensuing epigenetic silencing of proteasome genes
Article Snippet: U266 and RPMI 8226 cells were seeded in 24-well round-bottom plates (BD Biosciences) at a density of 3 × 10 5 cells/well and immediately transfected with 10 μM synthetic miR29b mimic (miR29b, Thermo Fisher Scientific #4464066) or miRNA mimic negative control (miR-NC, Thermo Fisher Scientific #4464058) using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #13778030).

Article Title: The circular RNA circNFIX regulates MEF2C expression in muscle satellite cells in spastic cerebral palsy.
Article Snippet: In addition, a miRNA mimic negative control (Thermo fisher, catalog #4464061) was included in the transfection along with the psiCHECK2-MEF2C-3’UTR_F1 to confirm that the observed effect in the previous experiments is indeed specific to hsa-miR-373-3p.

Article Title: Brain miR-137 governs growth and development via GH/IGF-1 signaling.
Article Snippet: Ten micrograms Mir-137 mimic (Cat.4464066, ThermoFisher) or miRNA mimic negative control (Cat.4464059, ThermoFisher) was dissolved in 2 μl saline for injection.

Article Title: The circular RNA circNFIX regulates MEF2C expression in muscle satellite cells in spastic cerebral palsy
Article Snippet: In addition, a miRNA mimic negative control (Thermo Fisher Scientific, catalog #4464061) was included in the transfection along with the psiCHECK2-MEF2C-3′UTR_F1 to confirm that the observed effect in the previous experiments is indeed specific to hsa-miR-373-3p.

Transfection:

Article Title: Distinct Effects of Respiratory Viral Infection Models on miR-149-5p, IL-6 and p63 Expression in BEAS-2B and A549 Epithelial Cells
Article Snippet: .. Cells at 70% confluency were transfected, as described before [ ], with miR-149-5p mimic, miRNA mimic negative control (mirVana, Invitrogen, Loughborough, UK), miR-149-5p inhibitor or miRNA inhibitor negative control (mirVana) at a total concentration of 5 nM using the Lipofectamine ® RNAiMAX Transfection Reagent (Invitrogen) in an Opti-MEM medium (Gibco). .. Epithelial cells were incubated with poly (I:C) (0.01–10 μg/mL for 24 and 48 h) or transfected with S1 or S2 (1 μg/mL) for 24 h. Cell viability was assessed by measuring lactate dehydrogenase (LDH) release [ ] at each timepoint using a SpectraMax iD5 Multi-Mode Microplate Reader (Molecular Devices, Wokingham, UK).

Article Title: Aberrant regulation of serine metabolism drives extracellular vesicle release and cancer progression.
Article Snippet: Article

Negative Control:

Article Title: Aberrant regulation of serine metabolism drives extracellular vesicle release and cancer progression.
Article Snippet: Article

Concentration Assay:

Image Search Results

a) Volcano plot representation of miRNA expression differences between microglia-derived VEVs and REVs following miRNA sequencing analysis; Upregulated genes (red) and down regulated genes (blue) with Log 2 foldchange>4 and Log 2 p value>4 for significance b) KEGG pathway analysis of miR-5099 associated target proteins determined via miRNA target analysis

Journal: bioRxiv

Article Title: Microglia-derived extracellular vesicles attenuate acute a-synuclein induced astrocyte inflammation

doi: 10.64898/2026.05.11.724371

Figure Lengend Snippet: a) Volcano plot representation of miRNA expression differences between microglia-derived VEVs and REVs following miRNA sequencing analysis; Upregulated genes (red) and down regulated genes (blue) with Log 2 foldchange>4 and Log 2 p value>4 for significance b) KEGG pathway analysis of miR-5099 associated target proteins determined via miRNA target analysis

Article Snippet: Cells were transfected 24 hours prior to additional treatments. miRNA mimics for mmu-miR-5099 (MED Chem express, Catalog# HY-R03245), mmu-miR-720 (Thermofisher, Catalog # 4427975) which has the same mature sequence as target Novel-miRNA-115 identified though miRNA sequencing, and scrambled miRNA (MCE, Catalog # HY-R04602 Lot#825593) were utilized for the transfections.

Techniques: Expressing, Derivative Assay, Sequencing

CircPTPN4 disrupts tight junction by upregulating ECE‐1 expression via miR‐145a‐5p sponging in BMECs. A) Quantitative RT‐PCR analyses of ECE‐1 mRNA levels in BMECs co‐cultured with Control neurons or Mg 2 ⁺‐free neurons, and transduced with either Control‐ShRNA or CircPTPN4‐ShRNA lentivirus ( n = 5; p < 0.0001, ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p = 0.0004, ShRNA‐CircPTPN4‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs). B) Western blot analyses of ECE‐1 protein levels in the cortex of Control and SE‐24 h mice infected with Control‐ShRNA or CircPTPN4‐ShRNA lentivirus. C) Bar graph quantifying ECE‐1 protein levels as the intensity ratio of ECE‐1 to GAPDH ( n = 5; p < 0.0001, ShRNA‐Ctrl‐treated SE‐24 h mice versus ShRNA‐Ctrl‐treated Ctrl mice; p < 0.0001, ShRNA‐CircPTPN4‐treated SE‐24 h mice versus ShRNA‐Ctrl‐treated SE‐24 h mice). D) Quantitative RT‐PCR analyses of ECE‐1 mRNA expression in Mg 2+ free N BMECs transduced with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐treated with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p < 0.0001, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0048, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p < 0.0001, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). E) Quantitative RT‐PCR analyses of ECE‐1 mRNA expression in Mg 2+ free N BMECs infected with OE‐Ctrl or OE‐CircPTPN4 lentivirus, and co‐treated with miR Control or miR‐145a‐5p mimics ( n = 5; p < 0.0001, miR Control and OE‐Ctrl BMECs versus miR‐145a‐5p mimics and OE‐Ctrl BMECs; p < 0.0001, miR Control and OE‐Ctrl BMECs versus miR Control and OE‐CircPTPN4 BMECs; p < 0.0001, miR Control and OE‐CircPTPN4 BMECs versus miR‐145a‐5p mimics and OE‐CircPTPN4 BMECs). F) Representative immunofluorescence images of Occludin staining in Mg 2+ free N BMECs infected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐infected with anti‐miR Control or miR‐145a‐5p inhibitors. G) Bar graph showing quantification of Occludin staining intensity in Mg 2+ free N BMECs infected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐infected with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p = 0.0066, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0021, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p = 0.0049, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). H) Bar graph showing the permeability of Mg 2+ free N BMECs transfected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐treated with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p = 0.0021, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0045, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p = 0.0002, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). I) Representative immunofluorescence images of Occludin staining in Mg 2+ free N BMECs co‐cultured with Control or Mg 2 ⁺‐free neurons, and infected with Control‐ShRNA or ECE‐1‐ShRNA lentivirus. J) Bar graph showing quantification of Occludin staining in Ctrl N BMECs or Mg 2 ⁺‐free N BMECs, and infected with Control‐ShRNA or ECE‐1‐ShRNA lentivirus ( n = 5; p < 0.0001, ShRNA‐ECE‐1‐treated Ctrl N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p =0 .0108, ShRNA‐ECE‐1‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs; p = 0.0004, ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p < 0.0001, ShRNA‐ECE‐1‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐ECE‐1‐treated Ctrl N BMECs). Values represent means ± S.E.M. Statistical analysis were performed using one‐way ANOVA followed by Tukey's test. Scale bar = 25 µm.

Journal: Advanced Science

Article Title: Circular RNA PTPN4 Contributes to Blood‐Brain Barrier Disruption during Early Epileptogenesis

doi: 10.1002/advs.202502250

Figure Lengend Snippet: CircPTPN4 disrupts tight junction by upregulating ECE‐1 expression via miR‐145a‐5p sponging in BMECs. A) Quantitative RT‐PCR analyses of ECE‐1 mRNA levels in BMECs co‐cultured with Control neurons or Mg 2 ⁺‐free neurons, and transduced with either Control‐ShRNA or CircPTPN4‐ShRNA lentivirus ( n = 5; p < 0.0001, ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p = 0.0004, ShRNA‐CircPTPN4‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs). B) Western blot analyses of ECE‐1 protein levels in the cortex of Control and SE‐24 h mice infected with Control‐ShRNA or CircPTPN4‐ShRNA lentivirus. C) Bar graph quantifying ECE‐1 protein levels as the intensity ratio of ECE‐1 to GAPDH ( n = 5; p < 0.0001, ShRNA‐Ctrl‐treated SE‐24 h mice versus ShRNA‐Ctrl‐treated Ctrl mice; p < 0.0001, ShRNA‐CircPTPN4‐treated SE‐24 h mice versus ShRNA‐Ctrl‐treated SE‐24 h mice). D) Quantitative RT‐PCR analyses of ECE‐1 mRNA expression in Mg 2+ free N BMECs transduced with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐treated with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p < 0.0001, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0048, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p < 0.0001, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). E) Quantitative RT‐PCR analyses of ECE‐1 mRNA expression in Mg 2+ free N BMECs infected with OE‐Ctrl or OE‐CircPTPN4 lentivirus, and co‐treated with miR Control or miR‐145a‐5p mimics ( n = 5; p < 0.0001, miR Control and OE‐Ctrl BMECs versus miR‐145a‐5p mimics and OE‐Ctrl BMECs; p < 0.0001, miR Control and OE‐Ctrl BMECs versus miR Control and OE‐CircPTPN4 BMECs; p < 0.0001, miR Control and OE‐CircPTPN4 BMECs versus miR‐145a‐5p mimics and OE‐CircPTPN4 BMECs). F) Representative immunofluorescence images of Occludin staining in Mg 2+ free N BMECs infected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐infected with anti‐miR Control or miR‐145a‐5p inhibitors. G) Bar graph showing quantification of Occludin staining intensity in Mg 2+ free N BMECs infected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐infected with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p = 0.0066, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0021, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p = 0.0049, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). H) Bar graph showing the permeability of Mg 2+ free N BMECs transfected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐treated with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p = 0.0021, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0045, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p = 0.0002, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). I) Representative immunofluorescence images of Occludin staining in Mg 2+ free N BMECs co‐cultured with Control or Mg 2 ⁺‐free neurons, and infected with Control‐ShRNA or ECE‐1‐ShRNA lentivirus. J) Bar graph showing quantification of Occludin staining in Ctrl N BMECs or Mg 2 ⁺‐free N BMECs, and infected with Control‐ShRNA or ECE‐1‐ShRNA lentivirus ( n = 5; p < 0.0001, ShRNA‐ECE‐1‐treated Ctrl N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p =0 .0108, ShRNA‐ECE‐1‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs; p = 0.0004, ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p < 0.0001, ShRNA‐ECE‐1‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐ECE‐1‐treated Ctrl N BMECs). Values represent means ± S.E.M. Statistical analysis were performed using one‐way ANOVA followed by Tukey's test. Scale bar = 25 µm.

Article Snippet: The miR‐145a‐5p mimics (HY‐ R00282 ), miRNA mimics negative Control (HY‐ R04602 ), miR‐145a‐5p inhibitor (HY‐RI00282), and miRNA inhibitor negative Control (HY‐RI04602) were purchased from Med Chem Express (MCE).

Techniques: Expressing, Quantitative RT-PCR, Cell Culture, Control, Transduction, shRNA, Western Blot, Infection, Immunofluorescence, Staining, Permeability, Transfection

Journal: Molecular Medicine Reports

Article Title: SLC7A1 , SGK1 and HMGB2 are overexpressed in cervical cancer tissues and the miR-23b-3p/HMGB2 axis regulates cell migration and invasion

doi: 10.3892/mmr.2025.13600

Figure Lengend Snippet: Target genes of miR-124-3p, miR-23b-3p and shared by both miRNAs. (A) The Venn diagram shows the target mRNAs of miR-124-3p and miR-23b-3p recorded in miRDB and TargetScan. (B) The table shows the name of the 136 genes predicted as targets shared by miR-124-3p and miR-23b-3p. miR, microRNA; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: As a negative control, mirVana miRNA Mimic Negative Control #1, Scrambled, (Invitrogen; Thermo Fisher Scientific, Inc.) was used.

Techniques:

a Schematic illustration of miRNA-responsive RNA OFF and ON switches coding the gene of interest (GOI). The OFF switch (left) inhibits translation of the protein of interest (POI) in the presence of miRNA activity, while the ON switch (right) promotes translation under the same conditions. b Schematic illustration of trans- protein splicing. In this reaction, split-inteins linked to their flanking peptides (exteins) ligate post-translationally, excising themselves and seamlessly joining the exteins to generate a mature protein. c Schematic illustration of “split ON switch,” a pair of ON switches coding protein fragments conjugated with split-inteins.

Journal: Nature Communications

Article Title: Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression

doi: 10.1038/s41467-025-60392-2

Figure Lengend Snippet: a Schematic illustration of miRNA-responsive RNA OFF and ON switches coding the gene of interest (GOI). The OFF switch (left) inhibits translation of the protein of interest (POI) in the presence of miRNA activity, while the ON switch (right) promotes translation under the same conditions. b Schematic illustration of trans- protein splicing. In this reaction, split-inteins linked to their flanking peptides (exteins) ligate post-translationally, excising themselves and seamlessly joining the exteins to generate a mature protein. c Schematic illustration of “split ON switch,” a pair of ON switches coding protein fragments conjugated with split-inteins.

Article Snippet: MiRNA mimics are small, chemically modified double-stranded RNAs that mimic endogenous miRNAs. mirVana miRNA Mimics (hsa-miR-21-5p, hsa-miR-302a-5p, and Negative Control #1) (Thermo Fisher Scientific, MC10206, MC12557, and 4464058) were used as mimic molecules in HEK293FT cells.

Techniques: Activity Assay

a Schematic illustration of the strategy to improve the ON/OFF ratio of ON switch systems. Introducing a “leak-canceller”, an OFF switch coding an inactive C-terminal fragment, together with the split ON switch, enables the suppression of leaky protein activity in miRNA-cells and enhances the ON/OFF ratio. b Relative hmAG1 intensity (hmAG1/iRFP670) of HEK293FT cells treated with miR-21-5p or negative control (NC) mimics. a.u., arbitrary units. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Welch’s t test, ** P < 0.01. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. c Representative microscopic images of HEK293FT cells transfected in the same conditions as shown in ( b ). Green fluorescence (left) and bright-field (right) images are shown for each condition. Scale bar, 200 µm. d Representative 2D flow cytometry plots. The horizontal axis shows the fluorescence intensity of iRFP670 (reference), and the vertical axis shows the fluorescence intensity of hmAG1 (reporter).

Journal: Nature Communications

Article Title: Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression

doi: 10.1038/s41467-025-60392-2

Figure Lengend Snippet: a Schematic illustration of the strategy to improve the ON/OFF ratio of ON switch systems. Introducing a “leak-canceller”, an OFF switch coding an inactive C-terminal fragment, together with the split ON switch, enables the suppression of leaky protein activity in miRNA-cells and enhances the ON/OFF ratio. b Relative hmAG1 intensity (hmAG1/iRFP670) of HEK293FT cells treated with miR-21-5p or negative control (NC) mimics. a.u., arbitrary units. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Welch’s t test, ** P < 0.01. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. c Representative microscopic images of HEK293FT cells transfected in the same conditions as shown in ( b ). Green fluorescence (left) and bright-field (right) images are shown for each condition. Scale bar, 200 µm. d Representative 2D flow cytometry plots. The horizontal axis shows the fluorescence intensity of iRFP670 (reference), and the vertical axis shows the fluorescence intensity of hmAG1 (reporter).

Techniques: Activity Assay, Negative Control, Transfection, Fluorescence, Flow Cytometry

a Schematic illustration of a toggle-like (two-output) system that displays hmAG1 fluorescence in the presence of target miRNA activity and iRFP670 fluorescence in its absence. “Normal toggle-like system” refers to a system composed of an ON switch coding full-length hmAG1 and an OFF switch coding full-length iRFP670, and “split toggle-like system” refers to a system composed of a pair of ON switches coding split hmAG1 fragments and a pair of OFF switches coding split iRFP670 fragments. b Normalised hmAG1 and iRFP670 fluorescence intensity of HEK293FT cells treated with various miR-21-5p mimic concentrations. Normalised hmAG1 intensity was calculated by normalising the intensity at 2 nM mimic concentration, and normalised iRFP670 intensity was calculated by normalising the intensity at 0 nM mimic concentration. Error bars represent means ± SD ( n = 3, biological replicate). Statistical analysis by two-sided Welch’s t test, * P < 0.05, ** P < 0.01. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. c Representative 2D flow cytometry plots of HEK293FT cells treated with various miR-21-5p mimic concentrations. The horizontal axis shows the fluorescence intensity of iRFP670 (reference), and the vertical axis shows the fluorescence intensity of hmAG1. d Euclidean distance between the centroid of the 0 nM mimic concentration plot and those of various mimic concentrations in the logarithmic 2D plot in ( c ). Error bars represent means ± SD ( n = 3, biological replicate). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression

doi: 10.1038/s41467-025-60392-2

Figure Lengend Snippet: a Schematic illustration of a toggle-like (two-output) system that displays hmAG1 fluorescence in the presence of target miRNA activity and iRFP670 fluorescence in its absence. “Normal toggle-like system” refers to a system composed of an ON switch coding full-length hmAG1 and an OFF switch coding full-length iRFP670, and “split toggle-like system” refers to a system composed of a pair of ON switches coding split hmAG1 fragments and a pair of OFF switches coding split iRFP670 fragments. b Normalised hmAG1 and iRFP670 fluorescence intensity of HEK293FT cells treated with various miR-21-5p mimic concentrations. Normalised hmAG1 intensity was calculated by normalising the intensity at 2 nM mimic concentration, and normalised iRFP670 intensity was calculated by normalising the intensity at 0 nM mimic concentration. Error bars represent means ± SD ( n = 3, biological replicate). Statistical analysis by two-sided Welch’s t test, * P < 0.05, ** P < 0.01. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. c Representative 2D flow cytometry plots of HEK293FT cells treated with various miR-21-5p mimic concentrations. The horizontal axis shows the fluorescence intensity of iRFP670 (reference), and the vertical axis shows the fluorescence intensity of hmAG1. d Euclidean distance between the centroid of the 0 nM mimic concentration plot and those of various mimic concentrations in the logarithmic 2D plot in ( c ). Error bars represent means ± SD ( n = 3, biological replicate). Source data are provided as a Source Data file.

Techniques: Fluorescence, Activity Assay, Concentration Assay, Flow Cytometry

a Schematic illustration of a two-input logic gate using split-intein, exemplifying an AND gate, in which an output protein is activated only in the presence of both of two different target miRNAs. b Four types of two-input logic circuits. Different colours correspond to those in the truth table, representative 2D flow cytometry plots, and bar graphs, respectively. MiR-21-5p and miR-302a-5p mimics were used as inputs. For example, the input pattern [10] coloured in blue means miR-21-5p was present while miR-302a-5p was absent. The representative 2D flow cytometry plot for each circuit is shown as an overlay of scatter plots for all four input patterns. The relative fluorescence intensity of hmAG1 (hmAG1/iRFP670) was normalised by the highest value for each circuit ([00] in NOR,[10] in A AND NOT B,[01] in NOT A AND B, and [11] in AND). Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Dunnett’s test for the ON state of each circuit, ***** P < 0.00001. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression

doi: 10.1038/s41467-025-60392-2

Figure Lengend Snippet: a Schematic illustration of a two-input logic gate using split-intein, exemplifying an AND gate, in which an output protein is activated only in the presence of both of two different target miRNAs. b Four types of two-input logic circuits. Different colours correspond to those in the truth table, representative 2D flow cytometry plots, and bar graphs, respectively. MiR-21-5p and miR-302a-5p mimics were used as inputs. For example, the input pattern [10] coloured in blue means miR-21-5p was present while miR-302a-5p was absent. The representative 2D flow cytometry plot for each circuit is shown as an overlay of scatter plots for all four input patterns. The relative fluorescence intensity of hmAG1 (hmAG1/iRFP670) was normalised by the highest value for each circuit ([00] in NOR,[10] in A AND NOT B,[01] in NOT A AND B, and [11] in AND). Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Dunnett’s test for the ON state of each circuit, ***** P < 0.00001. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file.

Techniques: Flow Cytometry, Fluorescence

a Schematic illustration of a two-type-input logic gate using split-intein, exemplifying a protein/miRNA-resonsive NOR gate, in which an output protein is activated only in the absence of both target protein and miRNA. b Two types of two-type-input logic circuits. L7Ae/miR-21-NOR gate exhibits hmAG1 fluorescence only in the absence of both L7Ae and miR-21-5p (upper). LIN28A/miR-302a-NOR gate exhibits hmAG1 fluorescence only in the absence of both LIN28A and miR-302a-5p (lower). Colours correspond to those in the bar graphs and representative 2D flow cytometry plots, respectively. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Dunnett’s test for the ON state of each circuit, ***** P < 0.00001. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. c Comparison of fold changes in cell type-dependent protein activity regulation among miR-302a-responsive OFF switch, LIN28A-responsive OFF switch, and LIN28A/miR-302a NOR gate. The fold changes in relative hmAG1 intensity were calculated as hmAG1/iRFP670 of HEK293FT cells divided by that of hiPSC. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Welch’s t test, * P < 0.05, ** P < 0.01. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. d Schematic illustration of a three-input logic gate using two orthogonal split-intein, exemplifying a three-input NOR gate, in which an output protein is activated only in the absence of all target miRNAs, and a three-input AND gate, in which an output protein is activated only in the presence of all target miRNAs. e Validation of miR-21/302a/206 NOR gate (left) and miR-21/302a/206 AND gate (right) in HEK293FT cell treated with different combinations of miR-21-5p, miR-302-5p, miR-206 mimics. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Dunnett’s test for the ON state of each circuit, ***** P < 0.00001. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression

doi: 10.1038/s41467-025-60392-2

Figure Lengend Snippet: a Schematic illustration of a two-type-input logic gate using split-intein, exemplifying a protein/miRNA-resonsive NOR gate, in which an output protein is activated only in the absence of both target protein and miRNA. b Two types of two-type-input logic circuits. L7Ae/miR-21-NOR gate exhibits hmAG1 fluorescence only in the absence of both L7Ae and miR-21-5p (upper). LIN28A/miR-302a-NOR gate exhibits hmAG1 fluorescence only in the absence of both LIN28A and miR-302a-5p (lower). Colours correspond to those in the bar graphs and representative 2D flow cytometry plots, respectively. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Dunnett’s test for the ON state of each circuit, ***** P < 0.00001. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. c Comparison of fold changes in cell type-dependent protein activity regulation among miR-302a-responsive OFF switch, LIN28A-responsive OFF switch, and LIN28A/miR-302a NOR gate. The fold changes in relative hmAG1 intensity were calculated as hmAG1/iRFP670 of HEK293FT cells divided by that of hiPSC. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Welch’s t test, * P < 0.05, ** P < 0.01. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file. d Schematic illustration of a three-input logic gate using two orthogonal split-intein, exemplifying a three-input NOR gate, in which an output protein is activated only in the absence of all target miRNAs, and a three-input AND gate, in which an output protein is activated only in the presence of all target miRNAs. e Validation of miR-21/302a/206 NOR gate (left) and miR-21/302a/206 AND gate (right) in HEK293FT cell treated with different combinations of miR-21-5p, miR-302-5p, miR-206 mimics. Error bars represent means ± SD ( n = 3), and data of each biological replicate are shown as a point. Statistical analysis by two-sided Dunnett’s test for the ON state of each circuit, ***** P < 0.00001. Each P -value is listed in Supplementary Table . Source data are provided as a Source Data file.

Techniques: Fluorescence, Flow Cytometry, Comparison, Activity Assay, Biomarker Discovery

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