No miRNA were found in Plasmodium and the ones identified in erythrocytes could not be correlated with infection.
BACKGROUND
The transcriptional regulation of Plasmodium during its complex life cycle requires sequential activation and/or repression of different genetic programmes. MicroRNAs (miRNAs) are a highly conserved class of non-coding RNAs that are important in regulating diverse cellular functions by sequence-specific inhibition of gene expression. What is know about double-stranded RNA-mediated gene silencing (RNAi) and posttranscriptional gene silencing (PTGS) in Plasmodium parasites entice us to speculate whether miRNAs can also function in Plasmodium-infected RBCs.


RESULTS
Of 132 small RNA sequences, no Plasmodium-specific miRNAs have been found. However, a human miRNA, miR-451, was highly expressed, comprising approximately one third of the total identified miRNAs. Further analysis of miR-451 expression and malaria infection showed no association between the accumulation of miR-451 in Plasmodium falciparum-iRBCs, the life cycle stage of P. falciparum in the erythrocyte, or of P. berghei in mice. Moreover, treatment with an antisense oligonucleotide to miR-451 had no significant effect on the growth of the erythrocytic-stage P. falciparum.


METHODS
Short RNAs from a mixed-stage of P. falciparum-iRBC were separated in a denaturing polyacrylamide gel and cloned into T vectors to create a cDNA library. Individual clones were then sequenced and further analysed by bioinformatics prediction to discover probable miRNAs in P. falciparum-iRBC. The association between miR-451 expression and the parasite were analysed by Northern blotting and antisense oligonucleotide (ASO) of miR-451.


CONCLUSIONS
These results contribute to eliminate the probability of miRNAs in P. falciparum. The absence of miRNA in P. falciparum could be correlated with absence of argonaute/dicer genes. In addition, the miR-451 accumulation in Plasmodium-infected RBCs is independent of parasite infection. Its accumulation might be only the residual of erythroid differentiation or a component to maintain the normal function of mature RBCs.
Xue
Xiangyang
X
Institute for Infectious Diseases and Vaccine Development, Tongji University College of Medicine, 1239 Siping Road, Shanghai 200092, China. wzxxy@yahoo.com.cn
Zhang
Qingfeng
Q
Huang
Yufu
Y
Feng
Le
L
Pan
Weiqing
W
D000818:Animals; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000469:parasitology*; D005260:Female; D015723:Gene Library; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D051379:Mice; D035683:MicroRNAs / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage; D010962:Plasmodium berghei / Q000235:genetics / Q000254:growth & development; D010963:Plasmodium falciparum / Q000235:genetics* / Q000302:isolation & purification; D016053:RNA, Protozoan / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; D016053:RNA, Protozoan