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Efficient Gene Targeting in Golden Syrian Hamstes by the CRISPR/Cas9 System
  • Z. Fan
  • W. Li
  • S. R. Lee
  • Q. Meng
  • T. D. Bunch
  • K. L. White
  • I. K. Kong
  • Zongde Wang, Utah State University
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The golden Syrian hamster is the model of choice or the only rodent model for studying many human diseases. However, the lack of gene targeting tools in hamsters severely limits their use in biomedical research. Here, we report the first successful application of the CRISPR/Cas9 system to efficiently conduct gene targeting in hamsters. We designed five synthetic single-guide RNAs (sgRNAs)—three for targeting the coding sequences for different functional domains of the hamster STAT2 protein, one for KCNQ1, and one for PPP1R12C—and demonstrated that the CRISPR/Cas9 system is highly efficient in introducing site-specific mutations in hamster somatic cells. We then developed unique pronuclear (PN) and cytoplasmic injection protocols in hamsters and produced STAT2 knockout (KO) hamsters by injecting the sgRNA/Cas9, either in the form of plasmid or mRNA, targeting exon 4 of hamster STAT2. Among the produced hamsters, 14.3% and 88.9% harbored germline-transmitted STAT2 mutations from plasmid and mRNA injection, respectively. Notably, 10.4% of the animals produced from mRNA injection were biallelically targeted. This is the first success in conducting site-specific gene targeting in hamsters and can serve as the foundation for developing other genetically engineered hamster models for human disease.
Citation Information
Fan Z., Li W., Lee SR., Meng Q., Bunch TD., White KL., Kong IK, Wang Z*. Efficient gene targeting in golden Syrian hamsters by the CRISPR/Cas9 system. PLOS One 2014 Oct 9;9(10):e109755. doi: 10.1371/journal.pone.0109755. *corresponding author