Fig 1: 5A2-DOT-X LNPs achieve CRISPR-Cas-based gene editing in therapeutic models.a To restore dystrophin expression, 5A2-DOT-10 LNPs encapsulating Cas9/sgDMD RNPs were injected into TA muscles of DMD exon 44 deletion mice weekly (three injections, 1 mg kg-1 sgDMD, n = 3). Three weeks after the last injection, TA muscles were collected to detect expression of dystrophin protein. b Immunofluorescence images indicated that 5A2-DOT-10 LNPs treatment successfully corrected dystrophin gene and restored the expression of dystrophin proteins in TA muscles. 5A2-DOT-10 LNPs nanoparticle only treatment was used as negative control (NC). Scale bar = 100 µm. Data was repeated two times independently with similar results. c Western blot analysis further confirmed the expression of dystrophin protein in the 5A2-DOT-10 LNPs encapsulating Cas9/sgDMD RNPs treatment group. 4.2% of dystrophin protein was restored. NC: 5A2-DOT-10 LNPs nanoparticle only; WT wild-type group. d To knockout the PCSK9 gene in mouse liver, 5A2-DOT-5 LNPs encapsulating Cas9/sgPCSK9 RNPs were administered to adult C57BL/6 mice via tail vein injection weekly (three injections, 2.5 mg kg-1 sgPCSK9, IV, n = 3). One week after the last injection, mouse serum and livers were collected for analyses. e The relative PCSK9 level in the serum was significantly decreased in 5A2-DOT-5 LNPs encapsulating Cas9/sgPCSK9 RNPs treatment group, detected using PCSK9 Elisa Kit. Data are presented as mean ± s.e.m. (n = 3 biologically independent animals). One-way ANOVA followed by Dunnett’s multiple comparison test was used to determine the significance of data. (*P < 0.05). f T7EI cleavage results from genomic DNA extracted from mice livers confirmed gene editing occurred at the PCSK9 locus. Red arrows indicate cleavage bands. Indel percentages shown under the gel image were measured by Sanger sequencing and TIDE analysis. Source data are in the Source Data file.
Supplier Page from Abcam for Mouse PCSK9 ELISA Kit