sgRNA design for DLT gene editing using CRISPR-Cas9 and in-silico mutation prediction in Rice cv. Hawara Bunar

Abstract The DWARF AND LOW TILLERRING ( DLT ) gene is a transcription factor for involved in Brassinosteroid (BR) biosynthesis. Manipulating BR biosynthesis will affect the height and tiller number of rice. CRISPR-Cas9 an accurate tool to edit sequence. accuracy site editing CRISPR-Cas9-mediated target determined by 20 nucleotide sequences sgRNA binding known as Protospacer Adjacent Motif (PAM). study aimed design predict mutation sites rice cv. Hawara Bunar. exon 1 was amplified using primer pair designed from reference gene. PCR product then sequenced, sequence used sgRNA. has located on targeted that corresponds Gras family protein domain exon1 were predicted be at through alignment amino acid It mutations change structure its function.