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Functional characterization of a surface antigen suppressing ApiAP2 in Eimeria tenella

ABSTRACT: Functional characterization of a surface antigen suppressing ApiAP2 in Eimeria tenella

PROVIDER: PRJNA1187613 | ENA |

REPOSITORIES: ENA

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Functional characterization of a surface antigen suppressing ApiAP2 in Eimeria tenella

Project description:Apicomplexa parasite Eimeria is the causative agent of coccidiosis, which leads to enteritis in animals and causes huge economic burden to the farm industry. The Apicomplexan Apetala2/ERF (ApiAP2) transcription factors have proved to play key roles in various processes in other Apicomplexa parasites. However, little is known about the function of ApiAP2s in Eimeria species. In this study, we functionally characterized an ApiAP2 by direct knockout mediated by CRISPR-Cas9. Our results showed that the invasion efficiency, total oocyst output and virulence to the host were significantly impaired after EtAP2-S1 depletion. RNA-Seq and CUT&Tag analyze showed that EtAP2-S1 targets on the promoters of numerous genes and its knockout results in upregulation of 59 sag genes. Additionally, the knockout strain exhibits significantly reduced virulence but provides excellent immune protection, which makes it a good vaccine candidate. This study demonstrates that EtAP2-S1 is a fitness coffering gene that suppress the expression of sag genes in E. tenella, and this is the first practice in developing gene knockout vaccine for the control of coccidiosis.

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Project description:In order to elucidate the infection mechanisms of Eimeria tenella and the chicken immune response, chickens were infected with Eimeria tenella strain Hougton sporozoites. Samples were taken at 0, 1, 2, 3, 4 and 10 days post-infection and mRNA sequenced. A dual-RNA seq analysis was carried out, comparing the expression of infected chickens during each sampling time point with uninfected chickens and comparing E. tenella samples during the infection with a sample of pure sporozoites. The results show a variety of response signals in the chicken, both previously known and unknown, as well as a clear role for a variety of infection-related genes in E. tenella

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Comparative analysis of Gallus gallus cecal epithelia following Eimeria tenella infection

Project description:Relative expression levels of mRNAs in chicken cecal epithelia experimentally infected with Eimeria tenella were measured at 4.5 days post-infection. Two weeks old chickens were uninfected (negative control) or were orally inoculated with sporulated oocysts of Eimeria tenella. Cecal epithelia samples were collected from >12 birds in infected or uninfected group at 4.5 d following infections, in which samples from 4 birds were pooled together to form a total 3 biological replicates in each group. Parasite merozoites were also collected from four infected chickens at 5 d after infections. Uninfected control samples, merozoites and infection group samples were selected for RNA extraction and hybridization on Affymetrix microarrays. We used Affymetrix GeneChip chicken genome arrays to detail the chicken cecal epithelia gene expression in the control and E. tenella-infected birds.

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