Coexistence of sense and anti-sense mRNAs of variant surface protein in Giardia lamblia trophozoites

Guo, Junli; Zheng, Wenyu; Wang, Yuehua; Li, Yao; Lu, Siqi; Feng, Xiao

doi:10.1016/j.bbrc.2014.01.074

Cited by 5 publications

(7 citation statements)

References 29 publications

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“…The strategy is based on the generation of dsRNA to silence expression of a selected gene by the RNAi pathway. Proteins characteristic of the RNAi pathway have been shown to be present and functional in this parasite [ 11 , 14 , 16 , 21 , 43 ], and it has been suggested that expression of some Giardia proteins, such as those of the VSP family that serve to evade the attack from the host immune system, are regulated by an RNAi-like mechanism [ 13 , 14 , 15 , 44 ]. Along this line, our data show that, indeed, it is possible to induce the RNAi machinery of Giardia to specifically attenuate the expression of selected genes.…”

Section: Discussionmentioning

confidence: 99%

“…Some studies have shown the presence of dsRNA or antisense RNA from endogenous ( α-Tub , RdRP , VSP ) or exogenous ( eGFP ) genes, which inhibit expression of these proteins; this is indicative of a gene-specific silencing mechanism in G . lamblia [ 15 , 45 ].…”

Section: Discussionmentioning

confidence: 99%

“…There are few studies employing RNAi in Giardia [ 11 , 13 , 14 , 15 ]. Several elements of the RNAi silencing pathway have been identified in Giardia , making conceivable its existence and therefore its potential usefulness to perform specific gene silencing.…”

Section: Introductionmentioning

confidence: 99%

“…Saraiya and Wang described that GlDcr is involved in the synthesis of microRNAs (miRNAs) derived from small nucleolar RNA (snoRNA), and that Ago of Giardia (GlAgo) participates in the miRNA-mediated silencing of the expression of proteins whose mRNA contain target sites for miRNA [ 16 ]. Recently, Guo et al proposed that the variant-specific surface protein ( VSP ) genes are transcribed simultaneously to sense and antisense strands, resulting in double-stranded RNAs (dsRNA) which are then degraded by a Dicer endonuclease, and that this regulation also involves antisense VSP transcription carried out by RdRP [ 15 ]. These results confirm that G .…”

Section: Introductionmentioning

confidence: 99%

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RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene

Marcial-Quino

Gómez-Manzo

Fierro

et al. 2017

Genes

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The microaerophilic protozoan Giardia lamblia is the agent causing giardiasis, an intestinal parasitosis of worldwide distribution. Different pharmacotherapies have been employed against giardiasis; however, side effects in the host and reports of drug resistant strains generate the need to develop new strategies that identify novel biological targets for drug design. To support this requirement, we have designed and evaluated a vector containing a cassette for the synthesis of double-stranded RNA (dsRNA), which can silence expression of a target gene through the RNA interference (RNAi) pathway. Small silencing RNAs were detected and quantified in transformants expressing dsRNA by a stem-loop RT-qPCR approach. The results showed that, in transformants expressing dsRNA of 100–200 base pairs, the level of NADHox mRNA was reduced by around 30%, concomitant with a decrease in enzyme activity and a reduction in the number of trophozoites with respect to the wild type strain, indicating that NADHox is indeed an important enzyme for Giardia viability. These results suggest that it is possible to induce the G. lamblia RNAi machinery for attenuating the expression of genes encoding proteins of interest. We propose that our silencing strategy can be used to identify new potential drug targets, knocking down genes encoding different structural proteins and enzymes from a wide variety of metabolic pathways.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene

Marcial-Quino

Gómez-Manzo

Fierro

et al. 2017

Genes

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“…Recently, it was reported that VSP genes are transcribed simultaneously in sense and antisense strands, which was proposed to result in the formation of double-stranded RNAs (dsRNAs) that are then degraded by the Dicer endonuclease complex [19]. Long dsRNAs can serve as templates for the biogenesis of small RNAs; this form of post-transcriptional gene silencing (PTGS) is a mechanism similar to the RNA interference (RNAi) pathway [20,21,22].…”

Section: Introductionmentioning

confidence: 99%

Stem-Loop RT-qPCR as an Efficient Tool for the Detection and Quantification of Small RNAs in Giardia lamblia

Marcial-Quino

Gómez-Manzo

Fierro

et al. 2016

Genes

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Stem-loop quantitative reverse transcription PCR (RT-qPCR) is a molecular technique used for identification and quantification of individual small RNAs in cells. In this work, we used a Universal ProbeLibrary (UPL)-based design to detect—in a rapid, sensitive, specific, and reproducible way—the small nucleolar RNA (snoRNA) GlsR17 and its derived miRNA (miR2) of Giardia lamblia using a stem-loop RT-qPCR approach. Both small RNAs could be isolated from both total RNA and small RNA samples. Identification of the two small RNAs was carried out by sequencing the PCR-amplified small RNA products upon ligation into the pJET1.2/blunt vector. GlsR17 is constitutively expressed during the 72 h cultures of trophozoites, while the mature miR2 is present in 2-fold higher abundance during the first 48 h than at 72 h. Because it has been suggested that miRNAs in G. lamblia have an important role in the regulation of gene expression, the use of the stem-loop RT-qPCR method could be valuable for the study of miRNAs of G. lamblia. This methodology will be a powerful tool for studying gene regulation in G. lamblia, and will help to better understand the features and functions of these regulatory molecules and how they work within the RNA interference (RNAi) pathway in G. lamblia.

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El arte de la evasión, fuente de nuevos fármacos

Fernández¹

2016

Medicina e Investigación

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Coexistence of sense and anti-sense mRNAs of variant surface protein in Giardia lamblia trophozoites

Cited by 5 publications

References 29 publications

RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene

RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene

Stem-Loop RT-qPCR as an Efficient Tool for the Detection and Quantification of Small RNAs in Giardia lamblia

El arte de la evasión, fuente de nuevos fármacos

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