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:: Volume 19, Issue 1 (spring 2021) ::
jmj 2021, 19(1): 10-20 Back to browse issues page
Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line
Behshad Montazeri-Najafabadi , Abbas Doosti * , Jafar Kiani
Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
Abstract:   (719 Views)
lncRNAs play their roles in cell growth, transcription, translation, and apoptosis. UCA1 is a type of lncRNA has a direct relationship with ovarian cancer. More than half of the women with ovarian cancer die within less than five years from the disease diagnosis. This study aims to knockout of UCA1 gene using the CRISPR/Cas9 technique in SK-OV-3 cell and studying its effect on cell factors involved in cancer progress and apoptosis.
Materials and Methods:
Two types of sgRNAs for UCA1 gene were designed. sgRNAs were cloned after synthesis in two vectors of PX459. The SK-OV-3 cell was transfected with two vectors carrying sgRNAs. The expression of FAS, BAK, BAX, and P53 pro-apoptosis genes and BCL2 and SURVIVIN anti-apoptosis genes were evaluated using the q-PCR. Besides, the cell proliferation rate and apoptosis induction were studied using MTT and flowcytometry.
The result showed the successful knockout of the UCA1 gene in the SK-OV-3 cell line. The expression of FAS, BAK, BAX, and P53 genes showed a meaningful increase related to the expression in manipulated cells compared to control cells (p˂0.05). Also, the expression decrease of BCL2 and SURVIVIN genes was seen (p˂0.05). MTT and flow cytometry results confirmed the cell proliferation decrease and the apoptosis increase.
UCA1 gene Knockout in the SK-OV-3 showed some positive effects in decreasing cancer cell proliferation and increasing the apoptosis rate can be said that inhibiting overexpression of the UCA1 gene will play a positive role in controlling the growth of ovarian cancer cells.
Keywords: UCA1, Ovarian cancer, CRISPR/Cas9
Full-Text [PDF 960 kb]   (200 Downloads)    
Type of Study: Research | Subject: daro sazi
Received: 2021/03/26 | Revised: 2021/05/17 | Accepted: 2021/05/5
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Research code: 1398/8
Ethics code: IR.IAU.SHK.REC.1399.017

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Montazeri-Najafabadi B, Doosti A, Kiani J. Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line. jmj. 2021; 19 (1) :10-20
URL: http://jmj.jums.ac.ir/article-1-1582-en.html

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Volume 19, Issue 1 (spring 2021) Back to browse issues page
فصلنامه دانشگاه علوم پزشکی جهرم Journal of Jahrom University of Medical Sciences
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