Fig. 5: Potential applications of RNA-based genetic circuits.
From: High-resolution and programmable RNA-IN and RNA-OUT genetic circuit in living mammalian cells
a The workflow to build RNA-sensing progesterone-producing cell factories in HEK293T cells. b The relative expression of STAR, CYP11A1 and HSD3B2 gene under different endogenous gene (RPS2, HSP90AB1, CANX) as input RNAs. c The biosynthesis pathway of progesterone and secreted progesterone levels in different RNA-IN and RNA-OUT genetic circuits. d Schematic of monitoring adipogenic differentiation of mesenchymal stem cells (MSCs). e Lipid droplet presence within the cells in brightfield and Oil Red O staining field, all experiments were repeated 2 times independently with similar results. f The microscopy images for the genetic circuits with or without crRNAs in adipogenic differentiation of MSCs. g The relative expression of adipocyte-related mRNA in undifferentiated and differentiated MSC cells. h The fluorescence of the genetic circuits with or without crRNAs targeted adipocyte-related mRNA in adipogenic differentiation of MSCs. i Schematic for monitoring trans-differentiation of HaCat from an epithelial-to-mesenchymal state. j Characterization of epithelial-to-mesenchymal transition (EMT) of keratinocytes (HaCat). Top: The cell morphology of HaCat with or without TGFβ factor induction. Bottom: The microscopy images for the genetic circuits with or without crRNAs in EMT induction of HaCat. k The relative expression of upregulated mRNAs of mesenchymal cell states in HaCat with or without TGFβ factor induction. l The fluorescence of the genetic circuits with or without crRNAs targeted mesenchymal-specific mRNAs with or without TGFβ factor induction in HaCat. m–p Sensing and killing two types of tumor cells with specific point mutations. Schematic of point-mutation-sensing-based tumor cell death (m). Point-mutation detection of endogenous TP53 R273H mutation (c.818G>A) in PANC1 tumor cells and Point-mutation detection of endogenous TP53 Y220C mutation (c.659A>G) in HuH7 tumor cells (n). Significance determined by two-tailed Student’s t test (Pleft = 0.004; Pright = 0.01). ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. Cell viability of HEK293T, PANC1, and HuH7 cells after transfection of different CASP sensor expressing HSV-tk (o, p). Non-targeting CASP sensor contains a scramble crRNA. Data are the average of three biological replicates ± s.e.m. Source data are provided as a Source Data file.
