AMB Volume 41, Issue 4, 2025 Pages 462-469 https://doi.org/10.59393/amb25410405
Inactivation of SNHG17 Using gRNA-Cas9 to Combat Helicobacter pylori-Associated Gastric Cancer
Dawood A.A., Al-Hadidi E.W.
Chronic Helicobacter pylori infections cause genetic changes and DNA damage in stomach epithelial cells. The gradual build-up of molecular defects that are a result of H. pylori infection within the stomach cells helps in the formation of stomach cancer with time. SNHG17 overexpression was related to low survival in gastric patients. The purpose of this work was to demonstrate how to build a CRISPR-Cas9 editing gene system to knockout SNHG17 by predicting a small guide RNA (gRNA) and linking it to the Cas9 enzyme. Several bioinformatics tools were employed to achieve the study’s goal, including the CRISPRDB, RNAfold, SimRNA 2.0, Geneious Prime, FUpred, HDOCK, and PyMol servers. gRNA93 and gRNA115 containing the target sequence AGGGA were selected. The cloning of gRNA-Cas9 was successfully loaded onto pCRII-TOPO. EcoRI is the most suitable type for cutting because it contains the required area for gRNA-Cas9, followed by NsiI and Mph1103I. E. coli BL21 is an appropriate bacterium for plasmid transformation. The lowest F-scores are reported between residues 65-75 and 1100-1130. The greatest docking score and RMSD for molecular docking are -2798.09 and 99.33, respectively. gRNA93 overlaps with the pocket of SNHG17. This study lays the groundwork for future research on gastric epithelial cell interactions with H. pylori, as well as the application of the gRNA-Cas9 system to reduce SNHG17 expression.
Keywords: Docking, gRNA, H. pylori, NONO, SNHG17
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