The scientists aimed to remove porcine OCT4 function as a model case using the CRISPR/Cas9 system. Injection of Cas9 and single-guide RNA (sgRNA) against OCT4 decreased the percentages of OCT4-positive embryos to 37-50% of total embryos,In addition, the combination of two sgRNAs led to large-scale deletion (about 1.8 kb) in the same chromosome. Next, they injected an enhanced green fluorescent protein (eGFP) vector targeting the OCT4 exon with Cas9 and sgRNA to create a knockin. They confirmed eGFP fluorescence in blastocysts in the inner cell mass. The CRISPR/CAS9 system provides a good tool for gene functional studies by deleting target genes in the pig.
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The scientists aimed to remove porcine OCT4 function as a model case using the CRISPR/Cas9 system. Injection of Cas9 and single-guide RNA (sgRNA) against OCT4 decreased the percentages of OCT4-positive embryos to 37-50% of total embryos,In addition, the combination of two sgRNAs led to large-scale deletion (about 1.8 kb) in the same chromosome. Next, they injected an enhanced green fluorescent protein (eGFP) vector targeting the OCT4 exon with Cas9 and sgRNA to create a knockin. They confirmed eGFP fluorescence in blastocysts in the inner cell mass. The CRISPR/CAS9 system provides a good tool for gene functional studies by deleting target genes in the pig.
www.geg-tech.com/Vectors