Editing of LINC00511 gene with a new CRISPR/Cas9 technique and evaluation of its effects on lung cancer cell line

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AZADBAKHT NARJES; DOOSTI ABBAS; Jami Mohammad Saeid

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Journal Paper

Paper Information

Journal: Journal of Jiroft University of Medical Sciences Year:2021 | Volume:8 | Issue:2 Page(s): 652-663

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Information Journal Paper

Title

Editing of LINC00511 gene with a new CRISPR/Cas9 technique and evaluation of its effects on lung cancer cell line

Author(s)

AZADBAKHT NARJES | DOOSTI ABBAS | Jami Mohammad Saeid | Issue Writer Certificate

Keywords

CRISPR-Cas9

Lung cancer

Non-coding RNAs

Apoptosis

Abstract

Introduction: Lung cancer is the second most common cancer in the world. The COR-L105 cell line isused as one of the most popular Lung cancer cell lines in various studies. Long Non-coding RNAs(lncRNAs) are a group of important cellular factors that play a key role in many processes such as growth, differentiation, replication, transcription, translation. LncRNA LINC00511 is a transcriptional regulatorand has been reported to increase its expression in various cancers. The aim of this study was to knockoutthe LINC00511 gene using a CRISPR/Cas9 in COR-L105 cells and to investigate its effects on cancerprogression and Apoptosis. Methods: In this experimental study, two sgRNAs were designed for LINC00511 gene and cloned into two CRISPR vectors, separately. Two sgRNAs vectors were transferred to COR-L105 cells usinglipofectamine. After confirmation of LINC00511 gene knockout from target cells, changes in cellproliferation and Apoptosis were assayed by MTT and flow cytometry. Furthermore, the expression ofgenes associated with Apoptosis and tumorigenesis was examined by real-time PCR. Results: LINC00511 gene knockout from COR-L105 cell line was performed. The expression of BCL2, survivin, EZH2, c-MYC, and MAX genes showed a meaningful decrease related to the expression in treated cells (edited) compared to the control cells (p˂ 0. 05). Increased expression of p21 and p57 geneswas also seen in the edited cells (p˂ 0. 05). Decreased cell proliferation and increased Apoptosis wereobserved in manipulated cells. Conclusion: Knocking of LINC00511 gene in Lung cancer cell line caused Apoptosis and decreased cellproliferation. Therefore, inhibiting the expression of the LINC00511 gene in cancer cells leads to thecontrol their proliferation.

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