In-silico Designing and Evaluation of gRNA for theCRISPR/Cas9 system against the Genes associated withTuberculosis

Tuberculosis caused by the bacterium Mycobacterium tuberculosis has been seen to show resistance against the prescribed drugs which are of uttermost importance and act as a first line of defense. The emergence of drug resistance has become a serious concern for the society and need for new and effective armamentarium persists. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated nuclease 9 (Crispr Cas9) is like a boon for the genetic engineering and have a wide application in gene editing. In the Mycobacterium tuberculosis, the gene Rv3378c is responsible for the production of a protein "1-TbAd" which helps the bacterium to survive inside the host for a period. The alteration in the gene Rv3378c can be potential therapeutics for the treatment of Tuberculosis. In the present study, a guide RNA (gRNA) was designed using CRISPOR tool to perform the necessary gene editing in the bacterium. The most suitable gRNA was selected with zero off targets. Zero off targets are necessary to decrease the chances of mistake in cutting the target sequence. This technique could serve as an important armamentarium to serve and save the people suffering from this disease.