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

Document Type : Short Communication

Authors

1 Department of Biology, Faculty of Basic Sciences, Qaemshahr Branch, Islamic Azad University, Mazandaran, Iran

2 Young Researchers and Elite Club, Qaemshahr Branch, Islamic Azad University, Mazandaran, Iran

3 Department of Medical Laboratory Sciences, Faculty of Medical Sciences, Gorgan Brach, Islamic Azad University, Golestan, Iran

4 ZistFanavari Novin Biotechnology Institute, Isfahan, Iran

Abstract

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 the
metastatic potential of a specific group of microRNAs called metastasis. Therefore, down-regulation of miRNAs at the
transcriptional level can reduce metastasis probability. The aim of this bioinformatics research is targeting of miRNAs
precursors using CRISPR-C2c2 (Cas13a) technique. The C2c2 (Cas13a) enzyme structure was downloaded from
the RCSB database, the sequence miRNAs and their precursors were collected from miRbase. The crRNAs were
designed and evaluated for their specificity by using CRISPR-RT server. The modeling 3D structure of the designed
crRNA was performed by RNAComposer server. Finally, HDOCK server was used to perform molecular docking to
evaluate 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 similarity
with the situation observed in normal and appropriate orientation was obtained. Despite high specificity, the correct
orientation 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 a
strong potential to inhibit metastasis. Therefore, crRNAs may be considered as an effective anticancer agent for further
research in drug development.

Keywords

Main Subjects

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