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Custom DNA/RNA Oligos> | The oligos of sgRNAs were synthesized (Genscript, China), and the complementary oligos were annealed to double-stranded DNA and ligated to the BbsI-digested pX330 vector (Addgene plasmid 423230) to generate the Cas9-sgRNA constructs, which were then transfected into PFFs using the Basic Fibroblast Nucleofection Kit (Amaxa Biosystems, Germany). | Get A Quote |
Oxysterol binding protein like 2 (OSBPL2), an important regulator in cellular lipid metabolism and transport, was identified as a novel deafness-causal gene in our previous work. To resemble the phenotypic features of OSBPL2 mutation in animal models and elucidate the potential genotype-phenotype associations, the OSBPL2-disrupted Bama miniature (BM) pig model was constructed using CRISPR/Cas9-mediated gene editing, somatic cell nuclear transfer (SCNT) and embryo transplantation approaches, and then subjected to phenotypic characterization of auditory function and serum lipid profiles. The OSBPL2-disrupted pigs displayed progressive hearing loss (HL) with degeneration/apoptosis of cochlea hair cells (... More