Fig. 3: Viral sense-and-destroy function in ALICE system.
From: Engineering antiviral immune-like systems for autonomous virus detection and inhibition in mice
a Schematic illustration of the design principle of ALICECas9+Ab system to autonomously sense-and-destroy of HSV-1. HSV-1 activates the ALICECas9+Ab system and ultimately initiates the expression of Cas9 and E317Ab under the control of the virus-inducible promotor (PALICE6). HSV-1-inducible Cas9 protein inhibits viral replication by deleting highly conserved sites on virus replication-related genes, while the expression of a recombinant monoclonal antibody that targets an epitope on glycoprotein D of HSV-1 (E317Ab) blocks viral infection. b qPCR analysis of EGFP-labeled HSV-1 mRNA levels in the ALICE system. White bars (Control), brown bars (ALICECas9), blue-gray bars (ALICEAb), red bars (ALICECas9+Ab) in (b, e, g). P values for all other groups versus Control group in the same gene. c The comparison study of antiviral effects between acyclovir and ALICE system in mammalian cells. P values for ALICECas9+Ab group versus Control group, and 50 μM ACV group on the same day. d Schematic for the protection function of ALICE system to host cells. Immune-like designer cells containing ALICE system were infected with EGFP-labeled HSV-1 (MOI = 1) for 3 h and then seeded on the inner chamber membrane of a Transwell® apparatus. Before this, HEK-293T cells had been seeded on the bottom of the outer chamber. EGFP expression of HEK-293T cells seeded on the bottom was profiled (e). f Schematic for the protection function of ALICE system to itself. HEK-293T cells were seeded on the bottom of the outer chamber and infected with EGFP-labeled HSV-1 (MOI = 1) for 3 h. Immune-like designer cells containing ALICE system were then seeded on the inner chamber membrane and embedded into the outer chamber. EGFP expression of immune-like designer cells containing ALICE system seeded on the inner chamber were profiled (g). The data in b, c, e, and g are expressed as means ± SD; n = 3 or 4 independent experiments in b, e, and g; n = 3 independent experiments in (c); P values were calculated by two-way ANOVA with Bonferroni’s post hoc test; n.s. not significant. Source data are provided as a Source Data file.
