RESUMO
RNA interference is a well established and widely used reverse genetic tool available for gene functional studies in trematodes. This technique requires the use of nonrelevant double-stranded RNA as control. However, several authors have reported inconsistencies associated with RNAi. We used RNASeq to analyze genes affected by nonspecific dsRNA exposure. We found only few genes presenting altered expression in schistosomula exposed to GFP or mCherry nonspecific-dsRNAs, most of them encoding uncharacterized proteins. Correlation analysis revealed that there are more differences among biological replicates, than due to treatment with nonspecific controls. These observations are of key relevance to other RNAi gene function assessment in other organisms.
Assuntos
Genes de Helmintos/fisiologia , Interferência de RNA , RNA de Cadeia Dupla/genética , RNA de Helmintos/genética , Schistosoma mansoni/genética , Animais , Biomphalaria/parasitologia , Expressão Gênica , Proteínas de Helminto/genética , Análise de Componente Principal , Esquistossomose mansoni/parasitologia , Esquistossomose mansoni/transmissão , Análise de Sequência de RNARESUMO
This study used qRT-PCR to examine variation in the expression of 13 myogenes during muscle development in four prenatal periods (21, 40, 70 and 90 days post-insemination) in commercial (the three-way Duroc, Landrace and Large-White cross) and local Piau pig breeds that differ in muscle mass. There was no variation in the expression of the CHD8, EID2B, HIF1AN, IKBKB, RSPO3, SOX7 and SUFU genes at the various prenatal ages or between breeds. The MAP2K1 and RBM24 genes showed similar expression between commercial and Piau pigs but greater expression (p < 0.05) in at least one prenatal period. Pair-wise comparisons of prenatal periods in each breed showed that only the CSRP3, LEF1, MRAS and MYOG genes had higher expression (p < 0.05) in at least one prenatal period in commercial and Piau pigs. Overall, these results identified the LEF1 gene as a primary candidate to account for differences in muscle mass between the pig breeds since activation of this gene may lead to greater myoblast fusion in the commercial breed compared to Piau pigs. Such fusion could explain the different muscularity between breeds in the postnatal periods.