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Iranian Journal of Basic Medical Sciences، جلد ۲۱، شماره ۲، صفحات ۱۸۱-۱۸۷
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عنوان انگلیسی CRISPR/Cas9, a new approach to successful knockdown of ABCB1/P-glycoprotein and reversal of chemosensitivity in human epithelial ovarian cancer cell line
چکیده انگلیسی مقاله Objective(s): Multidrug resistance (MDR) is a major obstacle in the successful chemotherapy of ovarian cancer. Inhibition of P-glycoprotein (P-gp), a member of ATP-binding cassette (ABC) transporters, is a well-known strategy to overcome MDR in cancer. The aim of this study was to investigate the efficiency and ability of CRISPR/Cas9 genome editing technology to knockdown ABCB1 gene expression in adriamycin resistant (A2780/ADR) ovarian cancer cell line and evaluate the sensitivity changes to doxorubicin. Materials and Methods: Three single-guide RNAs (sgRNAs) targeting the fourth and fifth exons of human ABCB1 gene were designed in this study. Expression level of ABCB1 was detected using quantitative real time PCR (qRT-PCR) after co-transfection of all three sgRNAs into A2780/ADR cell line and subsequent antibiotic selection. Drug sensitivity to doxorubicin was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results: The results showed that CRISPR/Cas9 system could significantly reduce the expression of P-gp. The dramatic decline in ABCB1 gene expression was associated with increased sensitivity of cells transfected with sgRNAs to doxorubicin. Conclusion: Based on the results of this study, it is concluded that the CRISPR-based systems, used in the present study, effectively down-regulated the target gene and acted as an ideal and cost-effective tool for gene editing of A2780/ADR cell line resulting in restoration of nonmalignant phenotype.
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نویسندگان مقاله لیلا نوروزی باروق | leyla norouzi barough
department of molecular medicine, school of medicine, qazvin university of medical sciences, qazvin, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی قزوین (Qazvin university of medical sciences)

محمدرضا ساروخانی | mohammad reza sarookhani
cellular and molecular research center, qazvin university of medical sciences, qazvin, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی قزوین (Qazvin university of medical sciences)

رسول صالحی | rasoul salehi
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

محمدرضا شریفی | mohammadreza sharifi
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

سحر مقبلی ن ژاد | sahar moghbelinejad
department of biochemistry and genetic, school of medicine, qazvin university of medical sciences, qazvin, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی قزوین (Qazvin university of medical sciences)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_10010.html
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article
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