In-silico Analysis for Prediction of Mutational Position and Designing of sgRNA for HDT701 Gene in Indian Rice cv. RPBio-226

Background: In eukaryotes, Histone acetylation and deacetylation plays a prominent role in controlling gene expression chromatin structure modifications. Both acetyltransferases (HAT) deacetylases (HDACs) work opposition to regulate acetylation. Reduced levels of histone H4 increased susceptibility the rice disease M. oryzae result from overexpression HDT701 rice. By changing defense-related pattern recognition receptor (PRR) genes rice, reduces activity innate immunity which promotes resistance oryzae. Crispr cas9 technology was created change modify characteristics, as well produce variety infections by focusing on possible biomolecules involved plant defense mechanisms. Therefore present study aimed design single guided RNA (sgRNA) predict mutational position HDT RPBio-226 cv. Methods: To begin with, DNA isolated using CTAB method. Specific pair primers designed reference for amplification gene. addition to, PCR product sequenced resulting sequence applied creation sgRNA. Furthermore, CHOPCHOP is Bioinformatic search tool used identify CRISPR–Cas guide targets. Result: The genomic quantified nanodrop found that concentration 800-1000ng/ul with purity 1.8. full amplified OsHDT701 sequenced. Based sequence, oligo generated http://chopchop.cbu.uib programme. target site designing sgRNA 168 basepair 190 deletion nucleotide at 174th position.