Golestan University of Medical Sciences Repository

The correction of ETV6/RUNX1 translocation in acute lymphocytic leukemia cells: a new gene targeting system by homologous recombination mechanism

Akbari, M. and Ebrahimabadi, S. and Golalipour, M. and Shahbazi, M. and Farazmandfar, T. (2020) The correction of ETV6/RUNX1 translocation in acute lymphocytic leukemia cells: a new gene targeting system by homologous recombination mechanism. Journal of Applied Genetics, 61 (1). pp. 67-73.

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Abstract

Regarding the uncertainty of the exact cause of the acute lymphocytic leukemia (ALL) caused by ETV6-RUNX1t(12;21) translocation, correcting genes of the ETV6 and RUNX1 in ETV6/RUNX1 fusion gene simultaneously on chromosome 12 may be effective in reducing leukemia malignancy. Thus, we designed an homologous recombination (HR) plasmid to target of the ETV6/RUNX1 fusion gene in the REH cell line containing the ETV6-RUNX1t(12;21) translocation. Cells were cultured and transfected by HR plasmid. The presence of the replacement cassette at specific location in the ETV6/RUNX1 fusion gene was verified by PCR and sequencing method. A quantitative gene expression assay was performed to evaluate changes in expression of ETV6, RUNX1, and ETV6/RUNX1 genes following editing. The cell viability was measured by trypan blue staining. The expression of the ETV6 gene was significantly increased in modified cells than unmodified cells by 10.9-fold. In contrast, the expression of the ETV6-RUNX1 fusion gene was significantly decreased in the modified cells compared with unmodified cells by 0.26-fold. The expression of the RUNX1 gene had no significant difference between modified and unmodified cells. The survival rate of edited cells was significantly decreased than unedited cells (p = 013). We designed a gene targeting system based on HR method to correct genes of ETV6 and RUNX1 simultaneously in ETV6/RUNX1 fusion gene on chromosome 12 containing ETV6-RUNX1t(12;21) translocation. The modification of this translocation may lead to reducing effects of the fusion gene�s damaging and the dosage compensation related to ETV6 and RUNX1 genes and subsequently reduce the effects of leukemia. This targeting system may open a window for treating leukemia as ex vivo. © 2019, Institute of Plant Genetics, Polish Academy of Sciences, Poznan.

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: genomic DNA; thymidine kinase; transcription factor ETV6; transcription factor RUNX1, acute lymphoblastic leukemia; acute lymphoid leukemia cell line; Article; cell culture; cell viability assay; DNA sequence; fusion gene; gene expression; gene expression regulation; gene overexpression; gene sequence; gene targeting; gene translocation; genetic transfection; homologous recombination; human; human cell; MTT assay; protein expression; real time polymerase chain reaction; RNA extraction; sequence alignment; survival rate; transcription factor ETV6 gene; transcription factor RUNX1 gene; unfolded protein response; virus isolation
Subjects: سیستم های خونی و لنفاوی WH
QU بیوشیمی
آسیب شناسی QZ
مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 14 Apr 2020 06:40
Last Modified: 14 Apr 2020 06:40
URI: http://eprints.goums.ac.ir/id/eprint/10513

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