Muhamad Mubarok

High Affinity K+ transporter 5 (HAK5) is one of the K + channel that involves in K + uptake. HAK5 gene expression is repressed under K + sufficient condition by the Auxin Response Factor 2 (ARF2) transcription factor. K + absorption can be increased by inactivation of ARF2 gene, e.g. by directed mutation using CRISPR/Cas9 method. The aims of this study were to create CRISPR/Cas9 construct carrying gRNA of OsARF2 gene and obtain transgenic rice Nipponbare containing CRISPR/Cas9- gRNAOsARF2. Two different sites in OsARF2 gene were succesfully designed as gRNA of the gene. Both oligo duplex RNA guides, gRNAOsARF2-A and gRNAOsARF2-B, were ligated into pDIRECT_21A plasmid using Golden Gate method. CRISPR/Cas9-gRNAOsARF2-A and CRISPR/Cas9-gRNAOsARF2-B cassettes were succesfully introduced separately into japonica rice cv. Nipponbare genome via Agrobacterium-mediated transformation, resulting putative transgenic lines. Analysis of Cas9 gene integration by PCR showed that 11 of 28 N-ARF-2-A...