Fig. 2: FUS-CRISPRee-mediated inducible suppression of endogenous genes.

a Schematic illustration of the FUS-CRISPRee system. b, c Representative flow cytometry data of CD81 (b) or CXCR4 (c) expression in FUS-CRISPRee-engineered Jurkat cells with gRNA targeting CD81 (b) or CXCR4 (c), or with non-targeting (NT) gRNA. The cells were stained with anti-CD81 (b) or anti-CXCR4 (c) antibodies four days after HS. d Relative CD81 mRNA expression 3 or 9 days after HS in cells in (b). P = 1.02 × 10⁻⁶ at 3 h in CD81 group. e, Relative CXCR4 mRNA expression in cells in (c). P = 2.99 × 10⁻⁷ at 3 h in CXCR4 group. f Percentage of CXCR4 + cells in Nalm6 cells engineered with CXCR4-targeting or NT FUS-CRISPRee with DNMT mutant with different treatments. g Kinetics of CXCR4 expression in cells engineered with CXCR4-targeting FUS-CRISPRee. h The migration ability (%) of the engineered FUS-CRISPRee Nalm6 cells in a transwell assay. In (b–h) HS, with 20 min HS; CT, without HS. In (f) FUS +; with 20 min FUS stimulation at 43 °C on cells in vitro. In d and (e), bar heights represent means of technical triplicates representative of two individual experiments. In (f and h), bar heights represent means of biological triplicates. Error bars represent s.e.m. Two-way ANOVA followed by Sidak’s multiple comparisons test was used in (d, e, and h). One-way ANOVA with multiple comparisons was used in (f). Source data are provided as a Source Data file. Created in BioRender. Liu, L. (2024) https://BioRender.com/h43m913.