Fig. 1

Preliminary design and functional validation of dCasRx translation enhancement system. A The eIF4GI was selected to fuse with dead CasRx. Then the CRISPR-dCasRx-eIF4GI increases translation of proteins based on sgRNAs by targeting sequences that bind specific RNAs. B The dCasRx-eIF4GI fusion protein, the NLS and sgRNA complementary DNA sequence of related genes were cloned into the plasmids containing CMV promoter and U6 promoter, respectively, for packaging into Lentiviral vectors. C To package into lentiviral vectors, the dCasRx-eIF4GI fusion protein and its complementary DNA sequence, sgRNA, were cloned into plasmids containing the CMV promoter and U6 promoter, respectively. D After constructing the no NLS CRISPR-dCasRx-eIF4GI tool targeting FTH1 (NNLS-DV-FTH1), the mRNA expression level of FTH1 were detected using qRT–PCR analysis. E The protein expression of FTH1 were verified using western blotting (WB) and the bar graph shows the relative expression level of FTH1. F After constructing the NLS CRISPR-dCasRx-eIF4GI tool targeting FTH1 (NLS-DV-FTH1), the mRNA expression level of FTH1 were detected using qRT–PCR analysis. G The protein expression of FTH1 were verified using WB and the bar graph shows the relative expression level of FTH1. Data are presented as the means ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and ns, no significant difference, represents P > 0.05