Journal of Cancer Research and Immuno-Oncology
Open Access

Abstract

A Systematic Biology Advances on sgRNA Design to Develop an Immunogenic Drug against Cancer Using CRISPR

Disha Patel, Khushal Patel*

In the field of genome editing, CRISPR/Cas9 is more suitable for clinical therapeutic applications, especially in the treatment of cancer. Here we wanted to design sgRNA that targets specific genes of helper T cells and cytotoxic T cells to alter them so that they can attack tumor cells. In other words, activating the immune system to treat cancer through the development of an immunogenic living drug. Among the main methods, the sgRNA of the target genes (PDCD-1, CTLA-4 and NYESO-1) was designed using the bioinformatics tool Benchling. The designed sgRNA was then cloned in silico into a lentivirus-based vector using the SnapGene gene designer. Isolation of an immunogenic drug can be done by transferring designed clones in vitro into the host and isolating pseudo-HIV viruses from the host, which consists of designed sgRNAs along with the CRISPR/Cas-9 machinery as a genome surrounded by an HIV viral envelope capable of binding T helper cells and cytotoxic T cells detected by protein-protein docking with PatchDock, followed by significantly improved results by FireDock and visualization in BIOVIA discovery studio. The sgRNAs were successfully engineered with good efficiency and potency to knock out PDCD-1 and CTLA-4 along with knock-in of NYESO-1. The developed immunogenic live drug has also shown positive results in binding efficiency to both T helper cells and cytotoxic T cells. Overall, this study showed that the key in silico results can be helpful in the development of immunogenic live drugs and will serve as a future aspect for in vitro treatment of cancer.

Published Date: 2025-01-15; Received Date: 2023-07-03