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. 2021 Aug 6;29(3-4):157–170. doi: 10.1038/s41434-021-00282-6

Fig. 13. COL7A1 gene exon 80 excision efficiency in RDEB keratinocytes transfected with CRISPR–Cas9 RNPs complexed to the HPAE-EB polymer.

Fig. 13

a PCR amplification from genomic DNA of a fragment spanning over the sgRNA target sites confirmed removal of exon 80 by the presence of a smaller molecular band (440 bp) in line with the distance between both cut sites (55 bp), marked with a lower arrow (red). Upper arrow (blue) indicates unedited DNA. Densitometry analysis from three independent experiments estimated an exon 80 excision of 43.2% using 4 µg of CRISPR–Cas9 RNP complex at 20:1 w/w ratio with HPAE-EB polymer (n = 3). b Collagen VII expression (red) was restored in RDEB keratinocytes after CRISPR–Cas9 RNP treatment, assessed by immunofluorescence staining, using DAPI (blue) as a nuclear stain. Magnification at 40×, scale bar 50 µm. Data were collected from two repeats of two independent experiments (n = 2).