Target-specific mutations efficiency at multiple loci of CRISPR/Cas9 system using one sgRNA in soybean

Jan 2, 2021Transgenic research

Efficiency of CRISPR/Cas9 gene editing at multiple sites in soybean using one guide RNA

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Abstract

Mutation frequencies of 64.71% for GmPDS, 60.0% for GmFAD2, and 42.86% for GmALS were achieved using a single sgRNA CRISPR/Cas9 system in soybean.

  • The CRISPR/Cas9 system was optimized to assess mutation efficiency in soybean's multiple-copy genes.
  • Simultaneous site-directed mutagenesis was successfully performed at two homoeologous loci with one sgRNA.
  • Chimeric and heterozygous mutations were the most common outcomes observed.
  • Phenotypic associations were identified with mutation patterns at specific target loci.
  • The findings indicate that while effective, the system exhibits relatively low transformation efficiency.

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