Journal: Nature Communications

Article Title: Muscle-specific gene editing therapy via mammalian fusogen-directed virus-like particles

doi: 10.1038/s41467-025-64200-9

Figure Lengend Snippet: a Schematic diagram illustrating the production of MuVLP Cas9 . b The levels of muscular fusogens, and Cas9 in VLPs, MuVLPs, VLP Cas9 , and MuVLP Cas9 . c ELISA quantification of Cas9 molecules per MuVLP Cas9 . The values and error bars represent the means ± s.d. of n = 3 technical replicates. d Scheme of MuVLP Cas9 or other controls cocultured with six different cell lines for detecting the specificity of MuVLP Cas9 for gene editing. e Immunofluorescence visualization of the Cas9 protein in cells cocultured with VLP Cas9 or MuVLP Cas9 . Red: anti-Cas9 antibody, green: F-actin labeled with Alexa Fluor 488 phalloidin, blue: nuclei labeled with DAPI. Scale bar, 20 μm. f Detection of exon 4 excision by genomic PCR in iC2C12 cells transfected with MuVLP Cas9 or other controls. The unedited genomic product is 883 bp long, and the gene-edited product (asterisk) is 445 bp long. g Sanger sequencing of amplicons confirmed the deletion of exon 4 and the generation of a fused intron 3/4 in genomic DNA from iC2C12 cells transfected with MuVLP Cas9 . The data are presented as the means ± s.d. of n = 3 biological replicates ( c ) or are representative of three independent experiments ( b , e – g ). Source data are provided as a Source Data file.

Article Snippet: Genomic DNA was extracted from tissues and cultured cells using a Genomic DNA Extraction Kit (D0063, Beyotime) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling, Transfection, Sequencing