Fig. 4 | Nature Communications

Fig. 4

From: Highly efficient RNA-guided base editing in rabbit

Fig. 4

Generation of a HGPS rabbit model using BE3 system. a The target sequence at the Lmna locus. The PAM- and sgRNA-target sequences are shown in green and black, respectively. The substituted bases are marked in red. Green and red lines indicate the normal and mutant mRNA splice forms, respectively. Mutant mRNA splicing results in a hypothetical 150-nucleotide deletion. b Alignments of mutant sequences from targeted deep sequencing. The targeted sequence is underlined. PAM site and substitutions are shown in green and red, respectively. The right column indicates the frequencies of the observed mutant alleles. WT wild-type. c Sanger sequencing chromatograms from WT and Lmna mutant rabbits (L5). The red arrow indicates the substituted nucleotide. Relevant codon identities at the target site are represented below the DNA sequence. d The predicted editing bar plot based on Sanger sequencing chromatograms from L5 by EditR. e Demonstration of the abnormal splice product using RT-PCR, showing a spliced product of 326 bp in mutant rabbits (L1 to L7) due to activation of the cryptic splice site. f Sanger sequencing chromatograms of the abnormal RT-PCR product confirmed the deletion of 150 nucleotides within exon 11 in the mutant rabbit (L5). g Photographs of WT (L8, 2.56 kg) and Lmna mutant (L5, 0.93 kg) rabbits at 3 months. h Radiograph and H&E staining of the skin from a 3-month-old Lmna mutant rabbit (L5) compared with a WT littermate (L8). The red triangle highlights the curvature of the spine. Red circles indicate subcutaneous adipose tissue. Scale bars: 200 μm

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