This technology involves editing the rice genome by introducing a guide RNA that recognizes a specific target sequence of the OsAAP2 amino acid transporter gene and a Cas endonuclease into rice cells, either as a vector or a ribonucleoprotein complex, followed by regeneration of the cells. The resulting edited rice and its seeds exhibit physicochemical starch properties distinct from those of the wild type.
As rice is a staple food for nearly half the world's population, improving starch quality—such as amylose content and gelatinization temperature, which determine taste and appearance—has become as important as increasing yield. While the roles of OsAAP3, OsAAP5, and OsAAP6 in yield and protein content have been reported, OsAAP2 has not been utilized for quality improvement. Furthermore, traditional GMO methods involving foreign gene insertion require significant time and cost for safety and environmental impact assessments.
This technology uses a CRISPR/Cas9 system targeting one of two sites (SEQ ID NO: 2 or 3) to induce mutations in OsAAP2. It also allows for the direct introduction of a ribonucleoprotein complex to avoid leaving behind foreign DNA, making the resulting mutations indistinguishable from natural ones. This approach can be applied to breeding new rice varieties with improved taste, customizing starch properties for processed rice, and securing genetic resources for rice quality research. It enables the rapid development of rice lines with enhanced taste and starch quality while bypassing the regulatory burdens associated with GMOs.
This invention was developed with support from the Ministry of Science and ICT for the functional analysis of genes regulating rice seed dormancy and pre-harvest sprouting resistance using the CRISPR/Cas9 system.
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