Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1

Manizheh Narimani,1 Mohammadreza Sharifi,2 Ali Jalili1 1Cancer and Immunology Research Center, Institute of Research for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran; 2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Scie...

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Autores principales:	Narimani M, Sharifi M, Jalili A
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	birc5 survivin crispr/cas9 nuclease aml kg1 cells hl60 cell Diseases of the blood and blood-forming organs RC633-647.5
Acceso en línea:	https://doaj.org/article/4ec3c036a8e5410e88b1c642b42dabb6
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spelling	oai:doaj.org-article:4ec3c036a8e5410e88b1c642b42dabb62021-12-02T02:27:33ZKnockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG11179-9889https://doaj.org/article/4ec3c036a8e5410e88b1c642b42dabb62019-11-01T00:00:00Zhttps://www.dovepress.com/knockout-of-birc5-gene-by-crisprcas9-induces-apoptosis-and-inhibits-ce-peer-reviewed-article-BLCTThttps://doaj.org/toc/1179-9889Manizheh Narimani,1 Mohammadreza Sharifi,2 Ali Jalili1 1Cancer and Immunology Research Center, Institute of Research for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran; 2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranCorrespondence: Ali JaliliCancer and Immunology Research Center, Institute of Research for Health Development, Kurdistan University of Medical Sciences, Sanandaj, IranTel +98-9183771862Email Ali130@gmail.comIntroduction: Human Baculoviral inhibitor of apoptosis repeat-containing 5 (BIRC5) which encodes survivin exhibits multiple biological activities, such as cell proliferation and apoptosis. Survivin is overexpressed in numerous malignant diseases including acute myeloid leukemia (AML). Recent studies have shown that the CRISPR/Cas9 nuclease-mediated gene-editing systems are suitable approach’s for editing or knocking out various genes including oncogenes.Methods and materials: We used CRISPR-Cas9 to knockout the BIRC5 in the human leukemic cell line, HL60, and KG1, and these cell lines were transfected with either the Cas9- and three sgRNAs expressing plasmids or negative control (scramble) using Lipofectamine 3000. The efficacy of the transfection was determined by quantitative reverse transcription-polymerase chain (RT-qPCR) and surveyor mutation assays. Cell proliferation and apoptosis were measured by MTT assay and flow cytometry, respectively.Results: We have successfully knocked out the BIRC5 gene in these leukemic cells and observed that the BIRC5-knocked out cells by CRISPR/Cas9 showed a significant decrease (30 folds) of survivin at mRNA levels. Moreover, cell death and apoptosis were significantly induced in BIRC5-CRISPR/Cas9-transfected cells compared to the scramble vector.Conclusion: We demonstrated for the first time that targeting BIRC5 by CRISPR/Cas9 technology is a suitable approach for the induction of apoptosis in leukemic cells. However, further studies targeting this gene in primary leukemic cells are required.Keywords: BIRC5, survivin, CRISPR/Cas9 nuclease, AML, KG1 cells, HL60 cellNarimani MSharifi MJalili ADove Medical Pressarticlebirc5survivincrispr/cas9 nucleaseamlkg1 cellshl60 cellDiseases of the blood and blood-forming organsRC633-647.5ENBlood and Lymphatic Cancer: Targets and Therapy, Vol Volume 9, Pp 53-61 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	birc5 survivin crispr/cas9 nuclease aml kg1 cells hl60 cell Diseases of the blood and blood-forming organs RC633-647.5
spellingShingle	birc5 survivin crispr/cas9 nuclease aml kg1 cells hl60 cell Diseases of the blood and blood-forming organs RC633-647.5 Narimani M Sharifi M Jalili A Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1
description	Manizheh Narimani,1 Mohammadreza Sharifi,2 Ali Jalili1 1Cancer and Immunology Research Center, Institute of Research for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran; 2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranCorrespondence: Ali JaliliCancer and Immunology Research Center, Institute of Research for Health Development, Kurdistan University of Medical Sciences, Sanandaj, IranTel +98-9183771862Email Ali130@gmail.comIntroduction: Human Baculoviral inhibitor of apoptosis repeat-containing 5 (BIRC5) which encodes survivin exhibits multiple biological activities, such as cell proliferation and apoptosis. Survivin is overexpressed in numerous malignant diseases including acute myeloid leukemia (AML). Recent studies have shown that the CRISPR/Cas9 nuclease-mediated gene-editing systems are suitable approach’s for editing or knocking out various genes including oncogenes.Methods and materials: We used CRISPR-Cas9 to knockout the BIRC5 in the human leukemic cell line, HL60, and KG1, and these cell lines were transfected with either the Cas9- and three sgRNAs expressing plasmids or negative control (scramble) using Lipofectamine 3000. The efficacy of the transfection was determined by quantitative reverse transcription-polymerase chain (RT-qPCR) and surveyor mutation assays. Cell proliferation and apoptosis were measured by MTT assay and flow cytometry, respectively.Results: We have successfully knocked out the BIRC5 gene in these leukemic cells and observed that the BIRC5-knocked out cells by CRISPR/Cas9 showed a significant decrease (30 folds) of survivin at mRNA levels. Moreover, cell death and apoptosis were significantly induced in BIRC5-CRISPR/Cas9-transfected cells compared to the scramble vector.Conclusion: We demonstrated for the first time that targeting BIRC5 by CRISPR/Cas9 technology is a suitable approach for the induction of apoptosis in leukemic cells. However, further studies targeting this gene in primary leukemic cells are required.Keywords: BIRC5, survivin, CRISPR/Cas9 nuclease, AML, KG1 cells, HL60 cell
format	article
author	Narimani M Sharifi M Jalili A
author_facet	Narimani M Sharifi M Jalili A
author_sort	Narimani M
title	Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1
title_short	Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1
title_full	Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1
title_fullStr	Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1
title_full_unstemmed	Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1
title_sort	knockout of birc5 gene by crispr/cas9 induces apoptosis and inhibits cell proliferation in leukemic cell lines, hl60 and kg1
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/4ec3c036a8e5410e88b1c642b42dabb6
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Knockout Of BIRC5 Gene By CRISPR/Cas9 Induces Apoptosis And Inhibits Cell Proliferation In Leukemic Cell Lines, HL60 And KG1

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