Design of crRNA to Regulate MicroRNAs Related to Metastasis in Colorectal Cancer Using CRISPR-C2c2 (Cas13a) Technique

Colorectal cancer (CRC) is the third most prevalent cancer with the second-highest mortality rate worldwide. microRNAs(miRNAs) of cancer-derived exosomes have shown promising diagnosis potential. Recent studies have shown themetastatic potential of a specific group of microRNAs called metastasis. Therefore, down-regulation of miRNAs at thetranscriptional level can reduce metastasis probability. The aim of this bioinformatics research is targeting of miRNAsprecursors using CRISPR-C2c2 (Cas13a) technique. The C2c2 (Cas13a) enzyme structure was downloaded fromthe RCSB database, the sequence miRNAs and their precursors were collected from miRbase. The crRNAs weredesigned and evaluated for their specificity by using CRISPR-RT server. The modeling 3D structure of the designedcrRNA was performed by RNAComposer server. Finally, HDOCK server was used to perform molecular docking toevaluate docked molecules' energy level and position. The crRNAs designed for miR-1280, miR-206, miR-195, miR-371a, miR-34a, miR-27a, miR-224, miR-99b, miR-877, miR-495 and miR-384 that showed high structural similaritywith the situation observed in normal and appropriate orientation was obtained. Despite high specificity, the correctorientation was not established in the case of crRNAs that designed to target miR-145, miR-378a, miR-199a, miR-320a and miR-543. The predicted interactions between crRNAs and Cas13a enzyme showed that crRNAs have astrong potential to inhibit metastasis. Therefore, crRNAs may be considered as an effective anticancer agent for furtherresearch in drug development.