• Gene therapy for human colorectal CD44+ cancer stem cells(CSCs) using CRISPR/Cas9 system
  • Seyed Taleb Houseini,1 Saeedeh saderi oskoiee,2 arash sattari,3 Mansoureh Azadeh,4 Roya BishehKolaei,5 Farkhondeh Nemati,6,*
    1. Department of Biology, Faculty of Basic sciences, Qaemshahr Branch, Islamic Azad University, Mazandaran, Iran
    2. Department of Medical science, Faculty of medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran
    3. Department of Medical laboratory sciences, Faculty of medical sciences, Gorgan Brach, Islamic Azad University, Gorgan, Iran
    4. ZistFanavari Novin Biotechnology Institute, Isfahan, Iran
    5. Department of Biology, Faculty of Basic sciences, Qaemshahr Bransch, Islamic Azad University, Mazandaran, Iran
    6. Department of Biology, Faculty of Basic sciences, Qaemshahr Bransch, Islamic Azad University, Mazandaran, Iran


  • Introduction: Colorectal CSCs have been therefore considered as a promising therapeutic target for colorectal cancer. The potential role of CRISPR/Cas9 in gene therapy has made it to become one of the hottest pots in cancer treatment. Different concepts of CRISPR/Cas9 mediated cancer therapy, including tumor related genes manipulating, tumor immunotherapy, tumor research modelling and anti-cancer drug resistance overcoming are established in various cancer types.
  • Methods: Geo dataset 100433 downloaded from Geo datasets. Genes are compared with logfc (fold change) for their expression in healthy and patients group and then sorted by their p-value. Also genes function and ontology are identified by DAVID database. Finally genes with most modification and related to critical pathway in immune system are selected and their best gRNAs(guide RNA) in CRISPR system are identified for editing by CHOPCHOP webtool.
  • Results: FOLR2 or folate receptor beta has hyperexpression among primary tumor patients. It plays an important roles in immune system and inflammatory response. The best gRNAs for this gene is identified through CHOPCHOP database.
  • Conclusion: Many experiences showed that immune genes play an important roles in cancer progression and metastasis, therefore they can be as a therapeutic target for CRISPR/Cas9 system and gene therapy.
  • Keywords: Colorectal cancer, CRISPR/Cas9, Gene therapy, gRNA