| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
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| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 34119684
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| MR4 number |
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| Parent parasite used to introduce the genetic modification |
| Rodent Malaria Parasite | P. berghei |
| Parent strain/line | P. berghei ANKA |
| Name parent line/clone |
Not applicable
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| Other information parent line | |
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| The mutant parasite was generated by |
| Name PI/Researcher | Yuda M, Nishi T |
| Name Group/Department | Department of Medical Zoology |
| Name Institute | Mie University School of Medicine |
| City | Mie, Tsu |
| Country | Japan |
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| Name of the mutant parasite |
| RMgm number | RMgm-5030 |
| Principal name | RvBL1-mNeonGreen |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | Not different from wild type |
| Gametocyte/Gamete | In fluorescence microscopy using mutant parasites expressing P. berghei RvBL1 fused with mNeonGreen, a strong expression was observed in male gametocytes. The expressed protein was distributed over the cytoplasm but was not detected in the nucleus. Signals of mNeonGreen were not detected in other intra-erythrocytic stages, including female gametocytes. |
| Fertilization and ookinete | Not tested |
| Oocyst | Not tested |
| Sporozoite | Not tested |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal (enhanced) GFP-tagged version of RvBL1 (RUVB1)
Protein (function)
In P. yoelii the CCR4-NOT complex is localized in cytoplasmic granules and is involved in translational regulation, playing important roles in development of male and female gametocytes. TRiC and the R2TP (Rvb1–Rvb2–Tah1–Pih1) complex play roles in protein synthesis as chaperones. TRiC is a molecular chaperone that is required for folding of proteins such as actin and tubulin. The R2TP complex is a co-chaperone of HSP90 that assists client proteins to form a large protein complex. Studies have demonstrated that the R2TP complex is essential for the preassembly of dynein arms, an essential step for cilium/flagellum formation.
Phenotype
In fluorescence microscopy using mutant parasites expressing P. berghei RvBL1 fused with mNeonGreen, a strong expression was observed in male gametocytes. The expressed protein was distributed over the cytoplasm but was not detected in the nucleus. Signals of mNeonGreen were not detected in other intra-erythrocytic stages, including female gametocytes.
Additional information
From the Abstract:
'Fluorescence microscopy showed that AP2-G expression was first observed in the parasite 12 h after erythrocyte invasion and peaked at 18 h when sexual features were first manifested in early gametocytes. Expression of AP2-G decreased with manifestation of sex-specific features. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) was performed at peak AP2-G expression and identified over 1000 binding sites in the genome. The main binding motif of the TF predicted from the binding sites was GTACNY. Predicted targets contained a number of genes related to protein biogenesis, suggesting that AP2-G plays a role in establishing a cellular basis required for sexual differentiation. AP2-G binding sites also existed upstream of gametocyte-specific TFs, namely AP2-G2, AP2-FG, and AP2-G itself. Furthermore, the target contained two AP2 TF-related genes. Disruption of these genes resulted in the arrest of ookinete development. These results suggest another role of AP2-G: activating a transcriptional cascade to promote conversion into early gametocytes.'
Other mutants |
*RMgm-5030
