Year-end Sale % OFF 
Abstract
ABSTRACTMalaria parasites have a complex life cycle with multiple developmental stages in mosquito and vertebrate hosts, and different developmental stages express unique sets of genes. Unexpectedly, many transcription factors (TFs) commonly found in eukaryotic organisms are absent in malaria parasites; instead, a family of genes encoding proteins similar to the plant Apetala2 (ApiAP2) transcription factors is expanded in the parasites. Several malaria ApiAP2 genes have been shown to play a critical role in parasite development; however, the functions of the majority of the ApiAP2 genes remain to be elucidated. In particular, no study on the Plasmodium yoelii ApiAP2 (PyApiAP2) gene family has been reported so far. This study systematically investigated the functional roles of PyApiAP2 genes in parasite development. Twenty-four of the 26 PyApiAP2 genes were selected for disruption, and 12 were successfully knocked out using the clustered regularly interspaced short palindromic repeat–CRISPR-associated protein 9 (CRISPR-Cas9) method. The effects of gene knockout (KO) on parasite development in mouse and mosquito stages were evaluated. Ten of 12 successfully disrupted genes, including two genes that have not been functionally characterized in any Plasmodium species previously, were shown to be critical for P. yoelii development of sexual and mosquito stages. Additionally, seven of the genes were labeled for protein expression analysis, revealing important information supporting their functions. This study represents the first systematic functional characterization of the P. yoelii ApiAP2 gene family and discovers important insights on the roles of the ApiAP2 genes in parasite development.
Highlights
Malaria parasites have a complex life cycle with multiple developmental stages in mosquito and vertebrate hosts, and different developmental stages express unique sets of genes
We attempted to disrupt 24 of the 26 ApiAP2 genes in the mouse malaria parasite P. yoelii using the clustered regularly interspaced short palindromic repeat–CRISPR-associated protein 9 (CRISPR-Cas9)-based methods we described previously [22], and we were able to knock out 12 of these 24 genes, including 3 genes homologous to genes that were not disrupted in P. berghei [9]
We carefully evaluated the morphologies of asexual/ sexual stages in ICR mice and sexual stages in Anopheles stephensi mosquitoes for the 12 gene KO mutants
Summary
Malaria parasites have a complex life cycle with multiple developmental stages in mosquito and vertebrate hosts, and different developmental stages express unique sets of genes. An ApiAP2 gene expressed in late oocysts and SG sporozoites was shown to be a major transcription factor that regulates gene expression in P. berghei’s sporozoite stage [17] Another P. berghei ApiAP2 gene, designated ap2-l, was found to play a critical role in the liver stage development of the parasite. We infected mice and mosquitoes with the mutant parasites to characterize phenotypic changes in parasite development, leading to functional characterization of two new P. yoelii ApiAP2 (PyApiAP2) genes that play important roles in gametocytogenesis and ookinete development, respectively. We successfully tagged seven PyApiAP2 proteins with sextuple hemagglutinin (6ϫHA) tags or the red fluorescent protein mCherry and revealed previously unknown protein expression profiles relevant to their functions in parasite development These results provide new insights into ApiAP2 gene functions and their roles in P. yoelii development in mosquito and vertebrate hosts
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
