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Volume 7, Issue 2 (Suppl)
J Ecosyst Ecography, an open access journal
ISSN:2157-7625
September 18-20, 2017
September 18-20, 2017 Toronto, Canada
Joint Conference
International Conference on
International Conference on
Environmental Microbiology and Microbial Ecology
&
Ecology and Ecosystems
J Ecosyst Ecography 2017, 7:2 (Suppl)
DOI: 10.4172/2157-7625-C1-030
The effect of double-stranded RNA on the expression of the homologous gene in
Toxoplasma gondii
Fatme Al Anouti
Zayed University, United Arab Emirates
T
he double-stranded RNA has been used in many organisms to interrupt gene expression at the post-transcriptional level.
We have explored the use of in vitro synthesized double-stranded RNA for gene expression study in
Toxoplasma gondii
. We
produced double-stranded RNAs homologous to the three well documented selectable markers the green fluorescent protein, the
uracil phosphoribosyl transferase and the hypoxanthine-xanthine-guanine phosphoribosyltransferase. Each dsRNA was efficiently
electroporated into the parasites and monitored for its effect on the expression of the homologous gene. The parasites electroporated
with the double-stranded RNA homologous to the green fluorescent protein exhibited reduced fluorescence for the green fluorescent
protein. The parasites electroporated with the double-stranded RNA homologous to the uracil phosphoribosyl transferase had low
enzymatic activity for the uracil phosphoribosyl transferase, while the parasites electroporated with the dsRNA homologous to the
hypoxanthine-xanthine-guanine phosphoribosyl transferase had low enzymatic activity for the hypoxanthine-xanthine-guanine
phosphoribosyl transferase. To investigate the in vivo longevity of the effects of the electroporated double-stranded RNA, we utilized
the uracil phosphoribosyl transferase. An operative uracil phosphoribosyl transferase assimilates 5-fluoro-2’-deoxyuridine ultimately
leading to parasite clearance. Parasites electroporated with the dsRNA homologous to the uracil phosphoribosyl transferase became
resistant to 5-fluoro-2´-deoxyuridine as a result of inhibited uracil phosphoribosyl transferase expression. Moreover, the effects of the
double-stranded RNA homologous to uracil phosphoribosyl transferase persisted for three successive propagations of the parasites.
Our study suggests that the double-stranded RNA could be a useful tool for gene silencing in
T. gondii.
Fatme.AlAnouti@zu.ac.ae