ABSTRACT

CRISPR/Cas9 exhibits the genetic modification or alteration or editing by nicking the desired genomic DNA at the target location. The accuracy modification depends on PAM (5'-NGG-3') sequence identified by Cas-9. The target sequence is 20 bases long on a crRNA array. The corrected PAM sequences to the target sequence are implanted in to a plasmid related to its accuracy and precision with no error in the genetic cut or modification. The given target sequence and PAM sequence are matched with the help of the software available like that of CRISTA and ccTOP. The amino acid sequence of the “Neu” (Herceptin) protein obtained from Expassy was subjected to CRISTA and ccTOP. The highest similarity of the SgRNA sequence showed a value of 0.98 to 0.99, counts up to 4 in number. Current manuscript an attempt to use software based correct identification of target sequence corresponding to the correct PAM sequence as recognized by Cas-9 for the generation of proper matched single-guided RNA sequence without the generation of the off targets formed either due to the repeated bulging of DNA. Furthermore, the characteristic properties of the Neu gene are also explored in order to assess the membrane position, polarity index, and hydropathy index. Thus the manuscript shall provide a glimpse of the software analysis for the corrected target sequence to the corresponding PAM sequence related to the precise genetic modification or alteration