EVALUATION OF KNOCKING DOWN OF THE RNAI MEDIATED GAMMA GLOBIN REPRESSOR INTO K562 CELLS, FOR GENE THERAPY OF BETA-THALASSEMIA

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ASGHARIAN ALI MOHAMMAD; BANAN MEHDI; NAJMABADI HOSSEIN

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Journal: Researcher Bulletin of Medical Sciences Year:2012 | Volume:16 | Issue:5 (83) Page(s): 234-240

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Title

EVALUATION OF KNOCKING DOWN OF THE RNAI MEDIATED GAMMA GLOBIN REPRESSOR INTO K562 CELLS, FOR GENE THERAPY OF BETA-THALASSEMIA

Author(s)

ASGHARIAN ALI MOHAMMAD | BANAN MEHDI | NAJMABADI HOSSEIN | Issue Writer Certificate

Keywords

BETA-THALASSEMIAQ3

TR4 SIRNAQ2

TR4 INHIBITIONQ2

GAMMA-GLOBINSQ2

K562 CELLSQ2

Abstract

Background and Aim: BETA-THALASSEMIA is a genetic disorder manifested by the presence of anemia in adult patients. One approach to treatment of BETA-THALASSEMIA is induction of the fetal gamma-globin gene. One of the gamma-globin repressors is a complex called DRED (Direct repeat erythroid-definitive). DRED is composed of TR2 and TR4 DNA binding subunits. The aim of this study was to set up the RNAi system to increase the expression of the gamma-globin gene.Materials and Methods: In this study, lipofectaminTM2000 was used to transfect siRNA molecules and pSV-b-Galactosidase vector was used as a reporter to monitor transfection efficiency. Real-time PCR method was used to measure TR4 knockdown and gamma-globin expression levels.Results: Our findings showed that K562 CELLS were transfected by 40% using Lipofectamine TM 2000. Also, the level of TR4 expression decreased by 44% after using TR4siRNA, even though the expression of gamma-globin gene was induced by 1.18 times.Conclusion: Despite a 44% knockdown of TR4, no increase in gamma-globin mRNA was observed. Two factors may count for this observation; first, TR4 knockdown may have been limited by our transfection efficiency. Second, simultaneous knockdown of TR2 and TR4 may lead to increased gamma-globin levels. In conclusion, although knocking down of TR4 expression occurred by using RNAi system, this can not be a solitary efficient way to induce the gamma-globin expression.

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APA: Copy

ASGHARIAN, ALI MOHAMMAD, BANAN, MEHDI, & NAJMABADI, HOSSEIN. (2012). EVALUATION OF KNOCKING DOWN OF THE RNAI MEDIATED GAMMA GLOBIN REPRESSOR INTO K562 CELLS, FOR GENE THERAPY OF BETA-THALASSEMIA. RESEARCHER BULLETIN OF MEDICAL SCIENCES (PEJOUHANDEH), 16(5 (83)), 234-240. SID. https://sid.ir/paper/18330/en

Vancouver: Copy

ASGHARIAN ALI MOHAMMAD, BANAN MEHDI, NAJMABADI HOSSEIN. EVALUATION OF KNOCKING DOWN OF THE RNAI MEDIATED GAMMA GLOBIN REPRESSOR INTO K562 CELLS, FOR GENE THERAPY OF BETA-THALASSEMIA. RESEARCHER BULLETIN OF MEDICAL SCIENCES (PEJOUHANDEH)[Internet]. 2012;16(5 (83)):234-240. Available from: https://sid.ir/paper/18330/en

IEEE: Copy

ALI MOHAMMAD ASGHARIAN, MEHDI BANAN, and HOSSEIN NAJMABADI, “EVALUATION OF KNOCKING DOWN OF THE RNAI MEDIATED GAMMA GLOBIN REPRESSOR INTO K562 CELLS, FOR GENE THERAPY OF BETA-THALASSEMIA,” RESEARCHER BULLETIN OF MEDICAL SCIENCES (PEJOUHANDEH), vol. 16, no. 5 (83), pp. 234–240, 2012, [Online]. Available: https://sid.ir/paper/18330/en

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