| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene disruption,
Introduction of a transgene
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| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 30154793
Reference 2 (PMID number) : 29515528
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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 |
P. berghei ANKA 676m1cl1 (RMgm-29)
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| Other information parent line | 676m1cl1 (RMgm-29) is a reference ANKA mutant line which expresses GFP-luciferase under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190). |
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| The mutant parasite was generated by |
| Name PI/Researcher | Fernandes P, Mueller AK |
| Name Group/Department | Centre for Infectious Diseases, Parasitology Unit |
| Name Institute | University Hospital Heidelberg |
| City | Heidelberg |
| Country | Germany |
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| Name of the mutant parasite |
| RMgm number | RMgm-4481 |
| Principal name | PbmaLS_05 (-) |
| Alternative name | PbmaLS_05 (−) GFP Luc(con) |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | Parasites lacking PbmaLS_05 showed a decreased ability to infect RBC (and immature reticulocytes), resulting in reduced peripheral parasite loads. Reduced mortality due to experimental cerebral malaria (ECM). None of the mice infected with mutant sporozoites developed ECM; in contrast, mice infected with WT sporozoites, displayed signs of ECM between day 7 and 9 p.i. Mice die from high parasitemia and anemia. |
| Gametocyte/Gamete | Not tested |
| Fertilization and ookinete | Not tested |
| Oocyst | Not tested |
| Sporozoite | No data provided. Assuming normal sporozoite production. |
| Liver stage | Quantification of both numbers and development through size measurements of in vitro exo-erythrocytic forms, suggested that deletion of PbmaLS_05 had no effect on the number or developmental size of intra-hepatic parasites, in vitro. A minor developmental delay was observed for KO parasites at 24 h post-invasion but not after 48 and 63 h.p.i.. Moreover, the branching of the apicoplast and segregation between daughter nuclei was also comparable between WT and KO intra-hepatic parasites, thus excluding any defect in apicoplast inheritance. Wild type parasite liver loads in vivo in mice. all KO-infected mice became patent with blood-stage infection on the same day as those infected with WT sporozoites, regardless of the route of infection used, indicating that deletion of PbmaLS_05 has no effect on both infectivity and intra-hepatic development. |
| Additional remarks phenotype | Mutant/mutation
The mutant lacks expression of PbmaLS_05 and expresses the fusion gene GFP-Luciferase under control of the constitutive eef1a promoter (in this study the gene PbmaLS_05 was also deleted in wild type P. berghei ANKA parasites).
Protein (function)
PbmaLS_05 has a predicted protein size of 306 kDa that contains two predicted transmembrane domains and one predicted P loop containing nucleoside triphosphate hydrolase domain.
The ortholog of PBANKA_140100 (PbmaLS_05), i.e., PF3D7_1302500, was initially identified as a putative antigen that is differentially expressed in intra-hepatic stages of P. falciparum radiation-attenuated sporozoites in comparison to Pf wild-type.
Evidence is presented for an apicoplast location.
Phenotype
Parasites lacking PbmaLS_05 showed a decreased ability to infect RBC (and immature reticulocytes), resulting in reduced peripheral parasite load. Reduced mortality due to experimental cerebral malaria (ECM). None of the mice infected with mutant sporozoites developed ECM; in contrast, mice infected with WT sporozoites, displayed signs of ECM between day 7 and 9 p.i. Mice die from high parasitemia and anemia.
No information is provided on mosquito stages. Assuming that wild type numbers of sporozoites are produced.
Quantification of both numbers and development through size measurements of in vitro exo-erythrocytic forms, suggested that deletion of PbmaLS_05 had no effect on the number or developmental size of intra-hepatic parasites, in vitro. A minor developmental delay was observed for KO parasites at 24 h post-invasion but not after 48 and 63 h.p.i.. Moreover, the branching of the apicoplast and segregation between daughter nuclei was also comparable between WT and KO intra-hepatic parasites, thus excluding any defect in apicoplast inheritance. Wild type parasite liver loads in vivo in mice. all KO-infected mice became patent with blood-stage infection on the same day as those infected with WT sporozoites, regardless of the route of infection used, indicating that deletion of PbmaLS_05 has no effect on both infectivity and intra-hepatic development.
Additional information
Expression throughout the complete life cycle (in all stages). Evidence presented for an apicoplast location. See also RMgm-4482 for a mutant expressing a C-terminal GFP-tagged version of PbmaLS_05.
Other mutants
See also RMgm-4482 for a mutant expressing a C-terminal GFP-tagged version of PbmaLS_05.
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*RMgm-4481
