Toxoplasma gondii: siRNA can mediate the suppression of adenosine kinase expression.
Adenosine kinase (AK) is one of the most important enzymes in the Toxoplasma gondii purine salvage pathway. Three siRNAs specific to the AK gene were designed in the present study. At 24h following electroporation, two of them (siRNA786 and siRNA1200) significantly reduced the mRNA level compared with mock electroporation (P <0.05). The ability to incorporate [3H]-adenosine in the parasites electroporated with 4 microM siRNA786 or 4 microM siRNA1200 was decreased to 39+/-11% and 39+/-7% of the mock electroporation, respectively. At the 48th hour of electroporation, the enzyme's activity was still significantly lower than that of mock electroporation. The data show the siRNAs transfected into cells can work efficiently to regulate gene expression in T. gondii. The application of siRNA in interrupting gene expression in T. gondii would be useful for elucidating gene function as a step toward development of anti-toxoplasmasis vaccines and therapeutic reagents.
Yu
Li
L
Institute of Clinical Pharmacology and Department of Pathobiology, Anhui Medical University, Hefei, Anhui 230032, PR China.
Gao
Yu-Feng
YF
Qiao
Zeng-Pei
ZP
Li
Cong-Lei
CL
Li
Xia
X
Shen
Ji-Long
JL
D000241:Adenosine / Q000378:metabolism; D000248:Adenosine Kinase / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics; D018274:Electroporation; D005347:Fibroblasts / Q000166:cytology / Q000469:parasitology; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D034622:RNA Interference / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014122:Toxoplasma / Q000201:enzymology* / Q000235:genetics*
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000248:Adenosine Kinase; D000241:Adenosine