SummaryRMgm-314
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 20019192 |
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 2.34 |
Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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The mutant parasite was generated by | |
Name PI/Researcher | R. Spaccapelo; C.J. Janse; J.B. Dame; A. Crisanti |
Name Group/Department | Department of Experimental Medicine |
Name Institute | University of Perugia |
City | Perugia |
Country | Italy |
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Name of the mutant parasite | |
RMgm number | RMgm-314 |
Principal name | Δpm4cl1; Δpm4cl6 |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Asexual blood stages show a slightly reduced growth rate and show a virulence-attenuated phenotype in mice (see 'Additional remarks phenotype'). |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | Oocyst production was reduced compared to wild type parasites |
Sporozoite | Oocyst production was reduced compared to wild type parasites. No sporozoite formation. |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation The mutant shows a virulence-attenuated phenotype in mice. Blood stages of the mutant are significantly less virulent than wild type P. berghei parasites. Mutant parasites failed to induce experimental cerebral malaria (ECM) in ECM-susceptible mice (i.e. C57Bl/6) and ECM-resistant mice (i.e. BALB/c) were able to clear infections. In vivo imaging of CD36-mediated sequestration of the schizont stage indicates that CD36-mediated sequestration is comparable to wild type (see also RMgm-32 for the method of in vivo imaging by measuring bioluminescence of the schizonts). In a paper published in 2024 (https://doi.org/10.1101/2023.10.24.563045) mutant parasites lacking expression of PM4 were analyzed during mosquito development. Oocyst production was reduced compared to wild type parasites and no sporozoites were produced. |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1034400 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1407800 | ||||||||||||||||||||||||
Gene product | plasmepsin IV | ||||||||||||||||||||||||
Gene product: Alternative name | PM4, plasmepsin 4 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) plasmid double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | SacII, PvuII | ||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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