Assessing the levels of cAMP response element binding protein1 (CREB1) after siRNA mediated knockdown in K562 cells

Authors
Zahra Deilami Khiabani 1
Mehdi Banan 2
Ali Mohammad Asgharian 3
Jalal Gharesouran 4
Hossein Najmabadi 5
1 Assistant Professor, Department of Microbiology, Faculty of Basic Sciences, Zanjan Branch of Islamic Azad University, Zanjan, Iran
2 Assistant Professor, Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran
3 Assistant Professor, Department of Biology, Faculty of Basic Sciences, Tonekabon Branch of Islamic Azad University, Mazandaran, Iran
4 Instructor, Department of Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
5 Professor, Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran
Abstract
Objective: CREB1 is an important downstream protein for many signaling pathways. By designing efficient siRNAs against CREB1, it may be possible to assess the role of molecules involved in signaling pathways in different cell types. In this research the efficiency of CREB1 knockdown by two different siRNAs in K562 cells has been studied.
Materials and Methods: siRNAs have been designed according to the criteria suggested by Reynolds et al. K562 cells were transfected by siRNA using Lipofectamine 2000. The efficiency of CREB knockdown has been assessed by quantitative relative Real-time PCR.
Results: Our results have shown that only one of the siRNAs has a high level of inhibitory effect on CREB1 gene expression. The expression of CREB1 by this siRNA was knocked-down by 87% in K562 cells.
Conclusion: In this research, although two siRNAs were designed according to the Reynolds et al. criteria, only one showed an inhibitory effect. Reasons other than the aforementioned criteria may be involved in effectiveness of siRNAs.

Keywords

Pathobiology Reserach
Volume 11, Issue 0
Spring 2008
Pages 49-55