Journal: Lab Animal

Article Title: Production of a heterozygous exon skipping model of common marmosets using gene-editing technology

doi: 10.1038/s41684-024-01424-0

Figure Lengend Snippet: a , Schematic representation of PSEN1 and TALEN used in the present study. The underlines indicate TALEN binding sites, the blue boxes indicate exons of the marmoset PSEN1 gene, and the red letters (AG) represent the 3′ splice site of intron 8. b , Left: the Surveyor assay of marmoset embryos after TALEN mRNA injection. +N: equal amount of embryo-derived PCR products and WT genomic DNA-derived PCR products were mixed for detecting the biallelic mutations of the PSEN1 gene; +/−: reagent control (the right two columns denote experimental controls included in the Surveyor mutation detection kit); M: GeneRuler 50 bp DNA Ladder (Thermo Fisher Scientific, SM0371). Right: the results of the sequencing analysis of subclones obtained from PCR products. c , Right: evaluation of exon skipping using TALEN mRNA-injected marmoset embryos. Left: illustration of the analysis. The black arrows represent the design positions of the primers used for RT–PCR. Samples with PCR product sizes smaller than 402 bp suggested exclusion of exon 9. M: 100 bp DNA ladder (New England Biolabs; N3231); RT (+): reverse transcriptase-treated samples; RT (−): samples with water added instead of reverse transcriptase.

Article Snippet: The Surveyor mutation detection kit (706025, Integrated DNA Technologies), a mutation analysis method that identifies mismatches in double-stranded DNA, was used to detect genetic modifications at the TALEN target site.

Techniques: Binding Assay, Injection, Derivative Assay, Control, Mutagenesis, Sequencing, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription