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.