Fig. 1: Schematic illustration of process of preparation of the Lenti@Cap-FAs and targeted delivery and M1 macrophages-specific RNA editing mediated by Lenti@Cap-FAs.
From: In vivo chemogenetic RNA editing of macrophages by bioengineered viruses for sepsis treatment
a A DNA construct encoding destabilized CasRx protein, which was fused by unstable dihydrofolate reductase (DHFR) domains, was constructed. The transgene construct is further installed with a macrophage-specific promoter. After the DNA construct is packed by lentivirus, a biocompatible layer of calcium phosphate (Cap) is applied to the outer surface to camouflage the virus. The Lenti@Caps was functionalized with folic acid (FA). TMP encapsulated by DSPE were prepared to stabilize dsCasRx and mediate target RNA knockdown by means of chemogenetic activation. b After the i.p. administration, the biomineralized lentivirus can target the inflammatory M1 macrophages, where the repeated administration of TMP@DSPE complexes can stabilize dsCasRx enzyme to knockdown NLRP3 mRNA. In the absence of TMP, structurally unstable dsCasRx is rapidly degraded via the ubiquitin-dependent proteasomal degradation route, and since other type cells do not express dsCasRx due to the macrophage-specific promoter, the system loses its ability to knockdown the target gene. Collectively, eliminating pathogens and control of inflammation through biomineralized and engineered lentivirus combined with TMP@DSPE mediated subset delivery represent an effective strategy for sepsis treatment. Created in BioRender. Yan, X. (2025) https://BioRender.com/feoajap.
