Fig. 5: Chemical-inducible HEAL systems (ChemHEAL) for regulation of endogenous gene transcription.

a Schematic of the ChemHEAL systems. In the presence of chemicals, specific fusion protein partners are recruited to each other, enabling the transactivator to localize to the gene of interest (GOI) and facilitate transcription of the endogenous gene with the assistance of the ΔdCas12f-sgRNA complex. b–e MCP and p65-HSF1 fragments were fused to chemical-inducible dimerization modules (ABI/PYL1 (a), NS3a/GNCR1 (b), NS3a/DNCR2 (c), or FRB/FRKB (d)). f–i Dose-dependent activation of HBG mediated by ChemHEAL system. HEK-293T cells transfected with plasmids encoding different ChemHEAL systems targeting HBG were treated with different concentrations of ABA (f), GZV (g), DNV (h), or Rapa (i) for 24 h. Relative mRNA levels of HBG were quantified using qPCR after induction. j–m Time-dependent activation of HBG mediated by ChemHEAL system. HEK-293T cells transfected with plasmids encoding different ChemHEAL systems targeting HBG were treated with 100 µM ABA (j), 10 µM GZV (k), 10 µM DNV (l), or 10 nM Rapa (m), and the relative mRNA levels of HBG were quantified by qPCR at different time points (0–48 h). n–q ChemHEAL-mediated activation of different endogenous genes. HEK-293T cells transfected with plasmids encoding different ChemHEAL systems targeting UCP1, ASCL1, or IFNγ were treated with 100 µM ABA (n), 10 µM GZV (o), 10 µM DNV (p), or 10 nM Rapa (q) for 24 h. Relative mRNA levels were quantified using qPCR after induction. Data in (f–q) are means ± s.d.; n = 3 independent experiments, with 3 technical replicates for each. Statistical significance was determined by two-tailed unpaired t test (f–q); ns, not significant, *P <  0.05, **P <  0.01, ***P <  0.001, ****P <  0.0001. Source data are provided as a Source Data file.