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Application of ZFN for Site Directed Mutagenesis of Rice SSIVa Gene

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Abstract

Many successful studies on genome editing in plants have been reported and one of the popular genome editing technology used in plants is Zinc Finger Nucleases (ZFN), which are chimeric proteins composed of synthetic zinc finger-based DNA binding domain and a DNA cleavage domain. The objective of this research was to utilize ZFNs to induce a double-stranded break in SSIVa, a soluble starch synthase involved in starch biosynthesis pathway, leading to the regulation of the SSIVa expression. The isoform SSIVa is not yet well studied, thus, by modifying the endogenous loci in SSIVa, we can explore on the specific roles of this gene in starch biosynthesis and other possible functions it might play. In this study, we used ZFN-mediated targeted gene disruption in the coding sequence of the SSIVa rice gene in an effort to elucidate the functional role of the gene. Generation of transgenic plants carrying premature stop codons and substitution events, revealed no SSIVa mRNA expression, low starch contents and dwarf phenotypes. Remarkably, based on our analysis SSIVa gene disruption had no effect on other starch synthesis related genes as their expression remained at wild type levels. Therefore, the engineered ZFNs can efficiently cleave and stimulate mutations at SSIVa locus in rice to

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Authors and Affiliations

  1. Department of Horticulture Life Science, Hankyong National University, Ansung, 17579, Korea

    Yu-Jin Jung & Kwon-Kyoo Kang

  2. Institute of Genetic Engineering, Hankyong National University, Ansung, 17579, Korea

    Yu-Jin Jung & Kwon-Kyoo Kang

  3. Department of Crop Science, Chungbuk National University, Cheongju, 28644, Korea

    Franz Marielle Nogoy & Yong-Gu Cho

  4. Department of Genetic Engineering, Kyunghee University, Yongin, 17104, Korea

    Sang-Kyu Lee

Authors
  1. Yu-Jin Jung
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  2. Franz Marielle Nogoy
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  3. Sang-Kyu Lee
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  4. Yong-Gu Cho
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  5. Kwon-Kyoo Kang
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Correspondence to Kwon-Kyoo Kang.

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Jung, YJ., Nogoy, F.M., Lee, SK. et al. Application of ZFN for Site Directed Mutagenesis of Rice SSIVa Gene. Biotechnol Bioproc E 23, 108–115 (2018). https://doi.org/10.1007/s12257-017-0420-9

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  • DOI: https://doi.org/10.1007/s12257-017-0420-9

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