Using RNAi to determine the function of Nopp44/46 in Trypanosoma brucei
Trypanosomiasis or "African Sleeping Sickness" is a Public Health concern for many countries in Sub-Saharan Africa. The parasite that causes African Trypanosomiasis, Trypansoma brucei, is transmitted to humans by the bite of an infected tsetse fly. Although, the pathology of Trypanosomiasis is not completely understood, studies have shown that T. brucei targets the blood stream of its victims and later spreads to various tissues. The parasite is fatal because it invades the central nervous system. Unfortunately, current treatments for African Sleeping Sickness are toxic, very expensive and inaccessible for those that most need them.
Nopp44/46 is a protein that has been found in the nucleolus of T. brucei. The exact function of Nopp 44/46 in trypanosomes is not known. It has been shown that Nopp 44/46 interacts with many proteins contained within the nucleolus and therefore it has been hypothesized that Nopp 44/46 may act as a scaffolding protein. In order to further determine the function of Nopp 44/46 within T. brucei the process of Post-Transcriptional Gene Silencing (PTGS) is being utilized.
The presence of double-stranded RNA in some cells has been known to hinder the expression of the corresponding gene. Scientists believe that this process is a natural defense mechanism to counter the invasions of viruses. A construct containing portions of Nopp 44/46 is being created for transfection into T. brucei. After inducing gene expression, the process of transcription will result in a "hair pin loop" RNA formation. The presence of the double stranded RNA should reduce expression of the endogenous Nopp44/46.Observing the cells lacking Nopp44/46 may help to elucidate the function of Nopp44/46 in T.brucei.
