Analyzing inhibition of BCL11A gene expression in K562 cells by RNAi
Loading...
Date
2013-07-23
Journal Title
Journal ISSN
Volume Title
Publisher
Plovdiv University Press “Paisii Hilendarski”
Abstract
RNA interference (RNAi), an effective approach to sequence-specific gene knockdown is widely used for the investigation of regulation of gene expression in various cells. BCL11A (B cell lymphoma 11A) plays a vital role in the evolutionarily different globin gene switches of mammals. In the current study, siRNA complementary to BCL11A was used to inhibit the BCL11A gene expression in erythroleukemic K562 cells and the expression was evaluated through real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis. On day 7 of cell culture, 1x106 K562 cells were transfected with lipofectamine containing BCL11A specific siRNA. GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) was used as the reference gene to confirm the relative expression level of BCL11A gene mRNA and BCL11A protein. After 48 h of transfection, BCL11A specific siRNA produced significantly reduction of BCL11A mRNA level in a dose-dependent manner. It also affects the level of BCL11A protein. BCL11A siRNAs were equally effective at reducing the expression level of BCL11A mRNA and protein.
Description
Keywords
BCL11A, K562, Thalassemia, RNAi, siRNA