Construction of Polycistronic tRNA-gRNA structures to study the function of some genes encoding ankyrin motif, related to rice panicle architecture, by using CRISPR/Cas9 technology

Abstract

The ankyrin repeat is one of the most common protein sequence motifs and has been detected in organisms ranging from viruses to human. Two genes OsANK1 and OsANK2 were determined implicated in rice panicle architecture through a GWAS analysis using a Vietnamese landrace panel of rice. The polycistronic tRNA-gRNA (PTG) structures that allow gene editing at mulitple sites, were built to characterize the function of OsANK1 and OsANK2, by using CRISPR/Cas9 technology. Two guide RNAs (gRNAs) were designed to cause gene mutation in the exon 4 region encoding ANK motif for each gene and were inserted into PTG structure using Golden Gate technology. At least one PTG structure was successfully obtained to edit OsANK1 and OsANK2 separetely or together.