Generation of Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting

2014-01-30


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Diagram of how gene-modified monkey generated

As published in Cell on January 30, 2014, Niu et al. reported precise gene targeting in cynomolgus monkeys by coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos and showed that this system enables simultaneous disruption of two target genes (Ppar-γ and Rag1) in one step, demonstrating that coinjection of one-cell-stage embryos with Cas9 mRNA and sgRNAs is an efficient and reliable approach for gene-modified cynomolgus monkey generation.

For details, refer to the paper:

Generation of Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos. Cell (2014) doi: 10.1016/j.cell.2014.01.027.

Niu et al.

Abstract:

Monkeys serve as important model species for studying human diseases and developing therapeutic strategies, yet the application of monkeys in biomedical researches has been significantly hindered by the difficulties in producing animals genetically modified at the desired target sites. Here, we first applied the CRISPR/Cas9 system, a versatile tool for editing the genes of different organisms, to target monkey genomes. By coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos, we successfully achieve precise gene targeting in cynomolgus monkeys. We also show that this system enables simultaneous disruption of two target genes (Ppar-γ and Rag1) in one step, and no off-target mutagenesis was detected by comprehensive analysis. Thus, coinjection of one-cell-stage embryos with Cas9 mRNA and sgRNAs is an efficient and reliable approach for gene-modified cynomolgus monkey generation.